AU2007201575A1 - System of hysteroscopic insemination - Google Patents

Description

11-04-2007 04:39PM FROM-A J PARK gton +64 4 4723358 T-168 P.004/099 F-266 -1-

AUSTRALIA

PATENTS ACT, 1990 COMPLETE SPECIFICATION FOR A STANDARD PATENT

ORIGINAL

Regulation 3.2 Name of Applicant: Actual Inventors: Address for service in Australix Invention Title: XY, INC.

MORRIS, Lee H. ALLEN, William and LINDSEY, Allison C., A J PARK, Level 11, 60 Marcus Clarke Street, Canberra ACT 2601, Australia System of hysteroscopic insemination The following statement is a full descaiption of this invention, including the best method of performing it known to us.

9447229,DOC COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:39PM FROM-A J PARK Vton +64 4 4T23358 T-168 P-005/092 F-266 wO V28311 PCTIS(11/02304 O WO 02128311 SYSTEM OF HYSTEROSCOPIC

INSEMINATION

This application claimsthebeneflt of and any applicable priority to, U.S. provisional application number 60/238,294, hereby inoorpoated by reference.

TECHNICAL FIELD This inventionrelates generally to the fieldofinleminilonofmammals. Specifically, it relates to systems to achieve inseminationwhichmay be particularly applicable once sperm o have been treated or processed in some manner, such as sorting the sperm through flow cytometry. MIore particularly, the invention may relate to systems to ahieve insemination, with a low number of spermatozoa as compared to conventional artificial insemination, thoughhysteroscopicinseminationtechniques. The inventionmaybeparticularly applicable to systems for inseminating equids, bovids and swine. Furthermore, the invention may be particularly applicable to sex selection of offspring in mammals.

BACKGROUND

Artificial insemination invarious forms, hasbeenusedto achieve succesful rates ofpregnancy and resulting offspring in mammals. Traditional forms ofAI include various in vivo technologies such as conventional intrauterine artificial insemnation techniques, ormore particularly, trans cervical artificial insemination. Natmal insemination doses may typically be large. For example, in some species, natural inseminationmay involve sperm numbers on the order of 2-20 x 10' sperm. In contrast, conventional AT techniques in the same species may require sperm numbers onthe order of 200-500 x 10f sperm. This can stillbe viewed as a large nmber of sperm, especially if the sperm are processed in some manner. Given the traditional requirements for high numbers of sperm to achieve acceptable numbers of pregnancies, other inseminationtechni queshavebeensoughtto address developing breeding technologies that may result in a lower sperm count for subsequent insemination. Low dose insemination has been particularly discussed in the PCT publication WO 99/33956, entitled "Sex-SpecifcInseminafionofllammls WithLowNumberofSperm Cells." Hysteroscopic insemination has been proposed and to some extent discussed in Morris et al., 2000, hereby incorporated by reference.

la COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:40PM FROM-A J PARK Vton +64 4 4T23358 T-168 P-006/099 F-266 WO 02/2311 PCT/S01/02304 0 WO2/28311 It should be preliminarily noted that all references mentioned in this application, and Z including additional reference not mentioned, are listed at the end of this written discussion, all of which should be considered as incorporated by reference.

High-speed sperm sorting using flow cytometry has been used successfully as a n 5 breeding technology to produce offspring in mammals, sach as horses, catte, sheep, rabbits, Spigs and humans. It can potentially be used for several other species as well. Technologies Shave beendeveloped to enhance ormodify the pregnancy and resulting offspring ofmammals, C particularly with regard to the processing of sperm and insemination techniques.

0 0 One potential limiting factor in optimizing the success of artificial insemination, as recognized in Pickett et al., 1989, may be inseminating with low numbers of spermatozoa- Pickett et al. recognizedthat apotential minimal recommended dosefor conventional artificial insemination in the mare may be as high as 500 x 10 6 progressively motile sperm. With low sort rates of around 700 spermatozoa per second in some sperm sorting technologies, it may take several days to obtainthe recommended dose of spermatozoa for artificial insemination.

This may not oly be impractical, but the viability oftbe spennatozoa may also be significanly reduced. Low-dose insemination techniques, herefore, may be a desirability to those skilled in the art to potentially reducethenumberofsperumatozoaneeded for acceptable fertility rates.

Additionally, what mightbe considered low doses for insemination may not typically result in acceptable fertility rates. Fertility rates may be considered acceptable or statistically comparable, for example, if they are achievable over a high sample size, range or percentage of the fertility rates resulting from conventional Al. Low-dose insemination techniques, therefore, may be a desirability to those skilledin the art to potentially reduce the number of spermatozoa needed to maximize fertility.

Whether in consideration of low-dose insemination or to improve AI fertility rates generally, an additional concern regarding artificial insemination is the efficiency of the procedure as a whole with regard to the resulting numbers of pregnancies. A number of 2 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:40PM FROM-A J PARK Vton 64 4 4723358 T-168 P-007/099 F-266 WO 02128311 PCTUS01/02304 procedural steps may have been used in conventional AI procedures, such as the synchronization of estrus in breeding meres; the preparation ofthe insemiation dose, more particularly the use of extendes for the dilution (Kenny et 1975) and resuspension of spermatozoa (using TALP or HEPES-bufered Tyrode's Medium, for example), centifugig the sample through a density or viscosity gradient (using PERCOLL or the like); assessing viability or motility; and the particulars of the insemination procedure. Insemination procedures have historically included ultrasound and rectally-guided techniques.

Hysteroscopic insemination of mares has also been conducted, as described in the Vasquez et al., 1998 and Manning et al, 1998 references. Only limited success, however, has been demonstrated in utilizing the above described procedures in conventional AI and in the Vasquez et al. and the Manning et al. procedures. In particular, neither reference may have produced statistically comparable conception rates to conventional AI. In particular, the Vasquez et al. and th Manning et al. references may have demonstrated conception rates of 33% for 3.8 x 106 spermatozoa and 22% for 1.0 x 106 spermatozoa, respectively, which may be considered non-comparable conception rates relative to conventional AI for the species involved. Despite the previous and substantial attempts at producing an efficientprocedure for AI, heretofore, a long felt but unsatisfied need for an efficient procedure for the hysteroscopic insemination ofmammals has xisted inbreedingtechnology. Furthermore, and given the traditional requirements for high numbers of spermatozoa to achieve acceptable numbers of viable pregnancies, as described above, a need for efficient low-dose hysteroscopic insemination has heretofore existed in current breeding technology.

Furthermore, deep intra uterine insemination of swine has been conducted, as described inthe Vasquez et al., 2000 reference. However, insemination was conducted with what might be considered a high number of spermatozoa, on the order of 20 x 10 7 spermatozoa. However, Vasquez et al., 2000, may have identified the long felt but unsatisfied need for an insemination procedure to address breeding technologies utilizing lower spermatozoa numbers for insemination, such as may result from current sperm sorting technologies.

Therefore, as may have been demonstrated from the Vasquez et al., 1998, Vasquez 3 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:40PM FROM-A J PARK ton 64 4 4723358 T-168 P-008/099 F-266 WO 02/28311 PCTUJSO1/02304 et al., 2000, and Manning et al., 1998, references, those skilled inthe art may have failedto address theidentifiedneed forproviding aniseminatontehniquepotetiallyresultinginhigh fertility rats iinammals and theresultinghighproduction ammals, higbfetility rats for low insemination dosages, and insemination techniqes to address issues of efficaiousness, particularly with regard to the hysteroscopic insemination technique.

One importantprocedural step withregardto inseinationprocedure, generally, isthe establishment of a insemination dose containing desirable numbers of viable and motile spermatozoa to potentially provide higher fertility rates. Procedures for the selection of motile spermatozoa may have been conducted with regard to conventional AI, for example in the reference Gnandahl et al., 1996 and in hysteroscopic insemination generally, by establishing adensity or viscosity gradient utilizing, for example, PERCOLL (Sigma Chemical Co., St. Louis, MO) alone or in combination with other substances. However, heretofore the substantial efforts to fractionate viable or motile sperm have not particularly addressed the identified needs for an insemination technique potentially providing high fertility rates, high fertility rates for low insemination dosages, and inseminationtechniquesto address issues of efficaciousness, particularly with regard to the hysteroscopic insemination technique.

A second potential issue with regard to insemination procedure, generally, is the establishment of a insemination dose containing desirable numbers of viable and motile spermatozoa to potentially provide higher fertility rates without aparticular motility test, as described above. The introduction of a density or viscosity gradient may introduce a stress to the spermatozoa that may actually reduce the actual number of viable and motile spermatozoa available from a particular sample. Heretofore, the substantial efforts to fractionate viable or motile sperm have not particularly addressed the identified needs for an insemination technique providing for high fertility rates, high fertility rates for low insemination dosages, and insemination techniques to address issues of efficaciousness, particularly with regard to the hysteroscopic insemination technique. In fact, due to the potential for reduction in the total number of viable or motile sperm from a particular inseminate sample previous attempts incorporating a density or viscosity gradient may have actually taught away from the present procedure of providing for an insemination technique 4 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:41PM FROM-A J PARK gton +64 4 4723358 T-168 P.009/099 F-266 WO 02/28311 PCT/US01/02304 Swhich may require less efforts to obtain a viable, low-dose insemination sample.

A third potential issue with regard to insemination procedure, generally, is the establishment of a compatible volume for the particular insemination technique. One recognized nee, as described above, is the desire to potentially provide higer fertility rates.

A second recognized need, also described above, is the ability to use low numbers of lC) spermatozoato potentially achieve highfertilityrates. Theinseminationdosevolume may be o determinedbytheparticularinseminationtechque. However,the dose voume may contain C a desirable number of spermatozoa to potentially provide for a higher rate of fertility.

SSubstantial attempts have been made to establish an appropriate inseminationtechlique that would allow for the appropriate number of spermatozoa, given the potential volume requirements of the insemation technique, to potentially provide acceptable fertility rates.

However, determining a compatible insemination dose volume for a particular insemination technique, to further potentially provide higher rates of fertility, has potentially not been established for hysteroscopicinsemination, as the reported fertility rates inVasquezet al. and Manning et al. may demonstrate.

Additionally, and as previously mentioned, the sperm sample may be processed prior to the insemination procedure. Conventional AI, for example, may provide for the use of extenders for the dilution (Kenny et al., 1975) and resuspension of spermatozoa. However, the particular media used may not be compatible with the insemination procedure itself Incompatibility ofthe sample media may result in lower deposition numbers of spermatozoa or dose volume or a lower fertility rate. Furthermore, the mode or form of the deposited insemination dose or the particular method of deposit during AI may further affectthe number of deposited spermatozoa available for conception. However, heretofore the substantial efforts directed toward insemination media may have not particularly addressed the need for providing an insemination technique potentially resulting in high frtlity rate, high fertility rates for low insemination dosages, and insemination techniques to address issues of efficaciousness, particularly with regard to the hysteroscopic insemination technique.

An additional factor to consider is the timing of insemination. Insemination timing COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:41PM FROM-A J PARK Von 64 4 4T23358 T-168 P-010/099 F-266 SWO 02/28311 PCT/US01/02304 0 N may be an important factor, for example, to sperm viability and longevity and the timing of t- the estrous cycle of the mammal. Particularly at issue might be the distant location of sperm sample acquisition the location of the male mammal) and ultimate location of the AI.

Previous efforts may have been made in conventional AI to preserve the sperm sample prior to insemination and to coordinate the insemination with the estrous cycle. However, tI heretofore the substantial efforts directed to insemination timing may have not particularly addressed the identified needs for, and may have even failed to understand the problems of o providing an insemination technique potentially resulting in high fertility rates, high fertility N rates for low insemination dosages, and insemination techniques to address issues of 0 10 efficaciousness, particularly with regard to the hysteroscopic insemination technique.

The source of the sperm sample may also be of importance to the resulting insemination. Epididymal acquisition ofthe sperm sample (obtaining sperm sample fromthe epididymis ofthe testis; ductules emerging posteriorly fom the testis thatholds sperm during maturation and that forms a tangled mass before uniting into a single coiled duct which is continuous with the vas deferens) may provide some inherent advantages as to timing of the insemination and viability of the sperm. However, heretofore the substantial efforts directed to sperm source, viability and inseminatontiming may have not fully addressed the identified needs for, and may have even failedto understand the problems of, providing an insemination technique potentially resulting in high fertility rates, high ferility rates for low insemination dosages, and insemination techniques to address issues of eflicaciousness, particularly with regard to the hysteroscopio insemination technique.

DISCLOSURE OF THE INVENTION The present inventors have recognized the potential problems associated with conventional methods of artificial insemination. Accordingly, embodiments of the present invention may provide for the production of a mammal through the use of artificial inseminationthat may address inadequacies ofprevious inseminationtecmiques and systems.

The invention may comprise, according to particular embodiments, a method ofproducing a mammal whereby poteniallyhighfrtility raes maybe accomplished, fertlityrates hichmay 6 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:42PM FROM-A J PARK VOR 64 4 4723358 T-168 P.011/099 F-266 SWO 02/28311 PCT/US01/02304 0

C

be statistically compatiblewithconventional AI results, and potentiallyhighfertilityrates with ,the use of low spermatozoa doses. More particularly, embodiments of the present invention Smay provide for the production of a mammal through the use ofhysteroscopic insemination techniques. Additionally, the present invention may comprise embodiments particularly directedatmammals suchas equids,bovids, and swie,mng othermammals. Embodiments ofthe present invention, therefore, may even be considered development away fromprevious In efforts of artificial insemination.

One object of the present invention, therefore, is to provide for the production of a 0 mammal utilizing an efficacious procedure. Therefore, a goal of the present invention is to provide atechnique of artficialinseminationfor mammal production such that lower umbers of spermatozoa may be used in the insemination dose relative to conventional AI and other insemnationtechniques, while, inpaticular embodiments oftheinvention, at least statistically comparable success rates in fertility are maintained.

Another object of the present invention is to provide for the production of a mammal utilizing an artificial inseminationprocedure that may potentially achieve high fertility rates consistent with lower spermatozoa production from breeding technologies such as sperm sorting. A goal of the present invention, therefore, is to provide a technique of artificial insemination for mammal production that achieves statistically comparable success rates in fertility, compared to conventional AI and other insemination techniques, with lower-doses of spermatozoa.

Furthermore, an object of the inventionis to provide for the production of amammal utilizing an artificial inseminationprocedure thatmay enhance steps involved inthe artificial insemination. To this end, one goal of the present invention is to provide a technique of artificial insemination for mammal production such that steps of estrous cycle timing, spermatozoasource, viability,longevity andprocessing, inseminationdosenmediand volume, and insemination timing may be optimized, particularly for low-dose insemination and potentially high fertility.

7 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:42PM FROM-A J PARK Vton +64 4 4T23358 T-168 P-012/099 F-266 WO 02/28311 PCTIUS01/02304 0 C An additional object ofthe presentinvention, according to preferred embodiments, is to provide fortheproduction ofaanmal throughhysteroscopic insemination. A goal ofthe present invention, therefore, is to provide a technique of artificial insemination for mammal production such that the insertion of the insemination dose, guiding of the insemination dose to the deposition site, deposition of the insemination dose at the appropriate location, in an appropriate mode or form, may be accomplished to achieve other objects and goals as previously stated. In accordance with particular embodiments of the invention, blister insemination andbubble or froth insemination may be introduced as preferred embodiments tooptimize fertility rates. A particular goal of the present invention is to provide atechniqae o 10 of artificial insemination for mammal production utilizing a catheter comprising a 0 videoendoscope for guiding and depositing the insemination dose.

Additionally, an object of the present invention, in accordance with particular embodiments, is to provide forthe productionofvarious mammal speciesutilizing anartificial insemination procedure. A goal ofthe present invention, therefore, is to provide a technique ofartificial insemination formarmamals such as equids, bovids and swine, among other species.

A finurther goal is to provide atechnique ofartificial inseminationfor various mammal species that additionally provides for low numbers of spennatozoa in the insemination dose and for potentially high fertility rates, particularly rates that may be statistically comparable to conventional AL.

Other objects ofthe vention are disclosedtbroughoutotherareas ofthe specificaton andclaims. In addition, the goals and objectivesmayapply either in dependentorindepedent fashion to a variety of other goals and objectives in a variety of embodiments.

BRIEF DESCRIPTION OF TIE DRAWINGS Figure 1 is a parially exploded and perspective diagrammatio view of the optical element, cannula, catheter and syringe in accordance with one embodiment of the present invention.

8 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:42PM FROM-A J PARK Vton 64 4 4723358 T-168 P.013/099 F-266 WO 02/28311 PCTUS01/02304

O

O

CN Figure 2 is a diagrammatic depiction of the reproductive organs of a female of a mammalian species and, in particular, a depiction of artificial insemination in accordance with one embodiment of the present invention.

Figure 3 is a magnified diagrammatic view ofan enshrouded insemination insertion embodiment of the present invention.

MODE(S) FOR CARRYING OUT THE PRESENT INVENTION o The basic concepts of the invention may be embodied in many different ways. The Sinventive conceptmay involve the materials, elements, apparatus, device and methods forthe production of a mammal through artificial insemination. Furthermore, while one preferred embodiment ofthe invention may be particularly directedto the productionofequids through artificial insemination, the broad concept ofthe invention shouldbe construed as a disclosure of the production ofmammals in general, and as indicated, to other mammal species such as bovids and swine.

As shouldbe understood, the present invention includes a variety ofaspects that may be used in various combinations depending upon the application's needs. The invention is intended to encompass a variety of embodiments of mammal production and combinations thereof. It involves both methods and devices to accomplish the various aspects explained.

In addition, while some methods and devices are disclosed, it should be understoodthatthese may be varied. Importantly, as to all ofthe foregoing all aspects should be understood to be encompassed by this patent both independently and in combination as set forth in the claims now or later issued.

Accordingly, one embodiment ofthe present invention may provide for the collection of spenn cells from amale of the species ofinterest. In accordance with ono embodiment, speam cells are collected from one or more stallions ofthe equine species. According to this embodiment, semen may be collected, and in preferred embodiments, semen may be collected with a commercially available artificial vagina, perhaps from at least one stallion of known 9 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:43PM FROM-A J PARK gton 64 4 4T23358 T-168 P-014/099 F-266 WO 02/28311 PCT/US01/02304 acceptable fertility. An artificial vagina such as one made available by Animal Reproduction Systems may be used with anin-line gel filter, and in prefered embodiments, used on alternate days throughoutcollection, After collection, the semenmay be evaluated forgel-free volume, motility, and sperm concentration. In accordance with another preferred embodiment, sperm cells may be collected from other male species ofmammal, particularly that ofbovids, equids or swine. An altrnative embodiment ofthepresent invention mayprovide for the collecting ofepididymal sperm cells obtained fomthe epididymis ofthetestis ofthemale species of the mammal. The alternative embodiment providing for the use of epididymal sperm may be incorporated with all other disclosed embodiments herein, either in single or in combination.

Furthermore, the present invention provides particular embodiments a hysteroscopic insemination sample comprising areservoir element a catheter system to whichthe reservoir elementisresponsive, andaplurality ofepididymal sperm cells contaiedwithinthe reservoir element.

After sperm collection, an artificial insemination sample may be established for the insemination of the female species. In accordance with one embodiment of the present invention, the sample may be prepared as having a low number of sperm compared to a natural insemination dosage for the mammal. The sample may have a low number of speua for particular breeding technologies, and in accordance with preferred embodiments, the sample may have a low number of sperm as the result of sorting the sperm for particular sexed sperm. In accordance with a preferred embodiment of the invention, the spermatozoa may be stained with Hoechst 33342 and sorted into X and Y chromosome-bearing populations based on DNA content using a commercially available SX MoFloe sperm sorter.

Additionally, an artificial insemination sample may be established at volumes, in accordance with preferred embodiments, at volumes between about 30 and 150 ul, less than about 500 ul, about 230 ul, and about 100 ul. One embodiment of the present invention is directed to establishing a hysteroscopic insemination compatible volume, preferably an insemination sample at a volume selected from a group consisting of: between about 30 and 150 ul, less than about 500 ul, about 230 ul, and about 100 ul. Furthennore, the present invention is directed to a hysteroscopic insemination sample comprising a reservoir element COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:43PM FROM-A J PARK Vton 64 4 4723358 T-168 P-015/099 F-266 WO 02/28311 PCTUSOI/023104 0 0 CN a catheter system to which the reservoir element is responsive, and a hysteroscopic compatible volume of sperm contained within said reservoir element. The hysteroscopic compatible volume of sperm contained witin said reservoir elementmay comprise avolume selected from a group consisting of: between about 30 and 150 ul less than about 500 ul, about 230 ul, and about 100 ul I Next, in accordance with preferred embodiments of the invention, the artificial insemination sample may be placed within a catheter or catheter system. In accordance with Salternative embodiments, thesamplemaybeplacedwithin aeservoirelementorother sample 0 holding element responsive to the catheter or catheter system. A catheter or catheter system should be understoodto define any device, system ormethod ofinsertion into canals, vessels, passageways, or body cavities to permit injection or withdrawal of fluids, and in accordance with preferred embodiments such injection or withdraw may provide the response of the reservoir element or sample holding element, or to keep a passage open.

Preferably, the catheter or catheter system may be used in conjunction with a guide element, and in preferred embodiments an optical element or device, and in preferred embodiments an illumination element, to provide guidance in the artificial insemination procedure, as more particularly described below. Preferred embodiments may provide the strobing of the illumination element. However, manual guidance may also be implemented.

In accordance with preferred embodiments of the invention, and as depicted in Figure 1, the insemination sample may be aspirated into an equine GIFT catheter (Cook Veterinary Products, Brisbane, Australia) using preferably a 6-mL disposable syringe attachedtothe injectionport on the distal end of the catheter. The loaded catheter may be withdrawn into a tube, or preferably an outer polypropylene cannula which may be responsive to an optical element and in accordance with preferred embodiments, passed down a working channel of a Pentax EPM 3000 videoendoscope (Pentax UK Ltd, Slough, Bucks

UK).

Accordingly, an instance, time or duration may be determined in which the artificial insemination may be appropriate. In accordance with preferred embodiments of the present 11 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:43PM FROM-A J PARK Vton +64 4 4723358 T-168 P-016/099 F-266 WO 02128311 PCT/USO1/02304 0 <N invention, a determination may be made as to an estrous time for a female mammal. In accordance with an embodiment of the invention, estras may be induced to determine the estrous time and, for multiple mares, even synchronized. Estrus may be defined as a state in Swbhich the female mammal is capable of conceiving and estrous cycle may be defined as the correlated phenomena of the endocrine and generative systems of a female mammal, In potentially from the beginning of one period of estus to not later than the beginning of the next. In accordance with a preferred embodiment, estrus may be induced, and for multiple o femnalemammals synchronized,by administeing a substance such as aprogestagen, preferably for mares altrenogest, and preferably 10ml orally, potentially for 10 consecutive days, O 10 followed by 250 tg cloprostenol im, potentially on day 11. Furthermore, a female mammal Smay be induced into ovulation atthetime ofinsemination Ovulationmaybe induced, andin preferred embodiments, by the administration of 3000 in human Chorionio Gonadotropin c(CG, Chorulon, Intervet, In., Millsboro, Holland), preferably administered intravenously at the time of insemination or up to approximately 8 hours previously. Ovulation may even be induced, in preferred embodiments, by the administration of from about 2000 to about 5000 in human Chorionic Gonadotropin.

In preferred embodiments, estrous cycles may be synchronized by administering a synthetic progestagen altrenogest (0.044mgkg Regumate; Hoechst Roussel Vet, Warren, New Jersey, USA) daily for 10 consecutive days. Luteolysis may be induced, in preferred embodimentswith the prostaglandin analogue, cloprostenol (250tgEstrunate, im.; Bayer Corporation, Agriculture Division, Shawnee Mission, Kansas, USA) administered on the eleventh day.

A time when the female mammal is appropriately fertile may be determined. The ovaries may be examined, andinpreferred embodiments examined ultrasonographically, and preferably every second day until afollicle, and in accordance with prefered embodiments a dominant follicle, preferably of> 30mm diameter, is detected- The female may be examined until a follicle, preferably pre-ovulatory, of preferably 35mm is detected. In one embodiment, the female may be inseminated during the same day as estrous inducement or synchronization, the same day as ovulation inducement or synchronization, or the same day 12 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:44PM FROM-A J PARK Vton +64 4 4723358 T-166 P-OIT/099 F-266 WO 02/28311 PCTIUS01o/2304 0 0 Sas estrus and insemination inducement or sychronization.

Next, in accordance withparticular embodiments ofthe invention, the guide element, Sand in preferred embodiments the optical element may be vaginally inserted into the female. Furthermore, the catheter and in preferred embodiments the reservoir element, may be inserted into the female. The sequence of insertion ofthe optical element (12) and the catheter (12) may be sequential or coincidental in time.

Cl The guide element or optical element (12) and the catheter may then be guided o through the vagina (20) of the female, as depicted in Figure 2. In accordance with one o embodiment, the optical element (12) and catheter may be manually guided. In fact, the catheter may be manually guided without the optical element However, accuracy in finding the UTJ and the potential result in increased fertility rates, particularly for low numbers of sperm and overall potential efficacy inthe procedure, may require amore accurate guidance procedure. Inpreferred embodiments, the optical element or endoscope, preferably aflexible endoscope, inpreferred embodiments having dimensions of 1.6 mlong with an outer diameter of 12 mm, may be guided through the cervix (22) and propelled forward through the uterine lumen or in additional embodiments, through a uterine horn of the female ofthe species. An added benefit of the use of the videoendoscope can be lack of a need to rectally guide the insemination process, as may have been required in past efforts by those of skill in the art.

The uterotubal junction (UTJ) (30) of the female mammal may then be located, preferably optically with the optical element The catheter may then be positioned in the vicinity of or proximate to the uterotubal junction. In accordance with preferred embodiments, the endoscope may be directedunder visual control along the uterine horn (26) ipslateral to the ovary containing the preovulatory follicle Prefexably the tip of the endoscope may be directed proximate to, and inpreferred embodiments within about of said uterotbal junction, and in preferred embodiments within about 3-5 cm ofthe papilla (32) of the uterotabal junction.

13 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:44PM FROM-A J PARK Cton 64 4 4T23358 T-168 P-018/099 F-266 WO 0228311 PCT/US01/02304 CN At least a portion of the artificial insemintion sample may then be extruded from the catheter In accordance with prferred embodiments, the outer cannula followed by the inner GIFT catheter containing the sperm suspension, may be extruded from the Sworking channel of the endoscope until the tip of the GIFT catheter touches the uterotubal junction, and in preferred embodiments, touches the papilla. In accordance with alternative embodiments of the present invention, a portion or at least a portion of the artificial insemination sample may be aspirated during extrusion from the catheter, therebypotentiaUy creating anaspirated sample. Suchaspirated samples shouldbe understoodto include bubbled Ssamples and frothed samples. An aspirated sample may not only provide better adherence to o 10 a surface in the vicinity of the UTJ, but may further allow for improved fertility rates.

0 Furthermore, an embodiment of the present invention provides ahysteroscopic insemination element comprising a uterus of a female species of a mammal, a plurality of sperm cells contained within said uterus, and an aspirated volume of media surrounding or interspersed with the sperm cells and to which the sperm cells are responsive. Such alternative embodiments should be construed as disclosed withregardto all embodiments ofthe present invention, either in single orin combination, and should be construedto be disclosed as such.

A low number of sperm may be deposited in the vicinity of the uterotubal junction.

In accordance with preferred embodiments, aplunger of the syringe may be depressed to deposit the sample, and in particular embodiments, a small volume (preferably perhaps -1 00p) of the sample, preferably onto the surface of the papilla. The guide element, and in preferred embodiments the optical element and the catheter may be withdrawn from the uterus ofthe female mammal. Inaccordancewithpreferredembodiment, filtered airmay be introduced within the uterus ofthe female to facilitate passage ofthe instruments through the uterine lumen. Furthermore, in accordance with preferred embodiments, the filtered air may be evacuated from the uterus, preferably simultaneous to the withdrawal of the optical element (12) and catheter The placing of a low number of sperm may be accomplished, in one preferred embodiment of the invention, by placing with the catheter a number of sperm, preferably mnnbers selected from: less than aboutten million sperm, less tan aboutfive million sperm, 14 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:44PM FROM-A J PARK Vton 64 4 4723358 T-168 P-019/099 F-266 WO 02/28311 PCTFIS01/02304 less than about two million sperm, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm.

Subsequently, the fertilization of an egg of the female may occur and the production of an offspring mammal from the fertilized egg. In accordance with preferred embodiments of the invention, success levels of fertilization may be statistically comparable to a conventional uterine body artificial insemination process. Statistically comparable success levels may be defined as fertilizationrates of at least about 75% success rates, at least about 65% success rates, at least about 60% success rates, at least about 50% success rates, at least about 45% success rates, and at least about 90% of a success rate practically expeiencedwith conventional AI for any particular species. Furtermore, success levels for the present invention may be statistically comparable to a conventional uterine body artifcial insemination process over a sample of cumulative fertilizations whichis greater than about 100, whichis greater than about 500, and which is greater than about 1000. Success rates may further statistically provide for at least a confidence level of at least about 95 percent confidence (potentially expressed as P> or equalto about 0.05), therefore potentially being statistically comparablewith conventional A.

Additionally, success rates for the present invention may potentially have the same P value over a range ofdiferig sper m numbers, potentially such as 1, 5 or 10 x 106 spe for equine.

Furthermore, a success level or rate may be expressed in temns of sample size, whereby the presentinventionmay provide, inpreferredembodiments, any ofthe aforementioned success rates over apower calculation ofatleast about 80 percent. Additionally, the success rates, in preferred embodiments, given particular insemination doses for sex-sorted stallion spermatozoa, may even apply to a low number of spermatozoa and may routinely produce fertility rates of at least about 90% of those rates resulting from conventional artificial insemination for aspecies. Additionally, according to particular embodiments ofthe present invention, the previous success rates may even be achieved for species of mammal such as bovids, equids, and swine.

Inregardto spermviability, longevity and mobility, for example, tepresentinvention may providefor establishinganartificial iseinationsampletilizingfreshsperm. Theterm COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:45PM FROM-A J PARK gton +64 4 4723358 T-168 P-020/099 F-266 SWO 02/28311 PCT/US01/02304 N "fresh sperm" may be broadly defined as sperm that has not been treated, processed or preserved in any manner such that the sperm viability, longevity and/or mobility might be compromised. Suchtreatment processing orpreservingmay include, for eample, the sorting of sperm, the freezing and subsequent thawing of spenm, the dilution and resuspension of sperm, and motility and viability testing or separation, generally, and in particular, Percoll gradient processing. The use of fiesh spermin accordance with embodiments of the present invention herein may permit the use of low numbers of sperm for insemination even, for Sexample, when using poor quality collected semen.

c o Alternative embodiments ofthepresent invention may broadly provide fortreatmnent, processing and preserving of insemination sperm. Alternative embodiments of the present invention may provide, for example, for the selection of the collected sperm cells more likely to achieve insemination. Accordingto one embodiment ofthepresentinveutionthe selection of desired sperm cells or sperm cells more likely to achieve insemination may comprise concentrating themore motile spermcollected. Additional embodiments may provide the step of cenrifuging the sperm through density gradients, and in preferred embodiments aPercoll gradient A potentially preferred embodiment may use a 90:45% Percoll gradient. An additional embodiment may comprise limiting the concentrationto less than about twice the starting concentration. Implementation of the Percoll gradient should be understood to encompass the use ofPercoll pre or postprocessing ofthe sperm, and inparticular, potentially pre or post sorting. Therefore, in accordance with embodiments herein, the Percoll gradient procedure may be used with "fresh sperm." The sample established from the desired or selected sperm cells may be used to establish the artificial insemination sample, potentially increasing the fertility rates due, at least in part, to the potentially higher rate of viability of the sample. The Percoll gradient procedure in accordance with the present invention may be conducted in conjunction wit h th e use oflower n mbers of sperm placed writbinthe depositing catheter, as more particularly described supra.

Additionally, establishing an artificial insemination sample may be provided by establishing asample having hysteroscopic compaSible media,ereby providing for potentially increased fertility rates and an efficacious insemination procedure. More particularly, an 16 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:45PM FROM-A J PARK gton 64 4 4723358 T-168 P-021/099 F-266 WO 02/28311 PCT/US01/02304 0 N embodiment of the invention may provide establishing an artificial insemination sample, and Sin preferred embodiments having a low number sperm compared to natural insemination doses, and providing for the establishment of an artificial insemination sample compatible media, for example extender, andinprefcedembodiments, askimmilk medim, such asEZ- Mixin CSTt (Animal Reproduction Systems, Chino, CA), preferably as a diluting media, potentially prior to additional processing. In accordance with preferred embodiments ofthe present invention, dilution may occurto no more tha a 2:1 ratio,to no more than a 5:1 ratio, and to no more than a 10:1 ratio, to potentially achieve at least a hysteroscopic compatible Svolume or media wit appropriate concentrations. Furthrmore, embodiments ofthe present inventionmay provide forthe use ofan extender, potentially a second extenderprovided after Ssperm processing andpotentially in conjunctionwithafirst extendr, as previously mentioned, to establish hbe sample utilizing hysteroscopic compatible media. The second extender may serve to resuspend the sperm sample after processing, and more particularly, provide for a sample utilizing hysteroscopic compatible media. According to preferred embodiments, the medium or second extender may comprise a TALP medium, a HEPES-buffered Tyrode's medium, and anAndrohepmedium. In accordance withpreleredembodiments ofthepresent invention,dilutionmaybeperformedwith askimmilkmedium suchasEZ-MixinCST, with aTALP medium, with aHE S-buffered Tyrode's medium, and with an Androhep medium, either single or in combination, to potentially achieve at least a hysteroscopic compatible volume or media with appropriate concentrations.

An additional embodiment ofthe present invention may also provide for an artificial insemination sample utilizing ahysteroscopic compatible media or mediumhaving acatheter coordinated viscosity. The viscosity may, for example, potentially facilitate the steps of extruding and depositing the sample. Furthermome in accordancewith potentially preferred embodiments ofthe present invention, the use ofcompatible media or medium may create an artificial insemination sample having a viscosity of preferably greater than about that of the blood of said mammal or greater than about that of a saline solution. Furthermore additional embodiments may provide for compatible media having a viscosity of greater than about 100cp, a media having a viscosity of greater than about 300cp, amediahaving a viscosity of greater than about 1000cp, a media having a viscosity of greater than about 300 0 cp, and a 17 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:46PM FROM-A J PARK Vton 64 4 4723358 T-168 P-022/099 F-266 WO 02/28311 PCT/US01/02304

O

C(N media having a viscosity of greater than about 6 0 0 0 op, each at about the mammal's average Sbody temperature.

SFurthermore, an embodiment of the present invention may provide a hysteroscopic inseminationsamplecomprising areervoirelement, a cathetersystemto which said reservoir element is responsive, aplurality of sperm cells containedwithin said reservoir element; and hysteroscopic compatible media contained within said reservoir element and to which said Ssperm cells are responsive. In accordance with additional embodiments, an insemination containment wherein saidplurality ofsperm cells are contaiaed within saidreservoir element o may include alow number of sperm cells as compared to the number of sperm cells typically found in a natural insemination.

Altering a property of the insemination specimen or sperm cell sample may be conducted according to the prsetinvention, and in accordance with preferred embodiments, determining an estrous time for a female of a species of said mammal and then altering a property of said insemination specimento establish an artificial insemination sample atabout said estrous time. Alternative embodiments may provide altering aproperty of the sample at about the time determined for the female of said species to be appropriately fertile, as previously defined. The present invention may also provide initiation of the altering of a property of the insemination specimen within a time selected from: within about twenty-four hours of said time determined for said female of said species of said mammal to be appropriately fertile, within about twelve hours of said time determined for said female ofsaid species of said mammal to be appropriately fertile, within about eight hours of said time determined for said female of said species of said mammal to be appropriately fertile, within about three hours of said time determined for said female of said species of said mammal to be appropriately fertile, and within about one hour of said time determined for said female of said species of said mammal to be appropriately fertile. This alteration may consist of preparing the sample, sorting the sperm, thawing the sperm, or the like.

One particular embodiment of the present invention may provide for esablishing a hysteroscopic compatible media and the concentration of sperm to select sperm more likely to achieve insemination. Particularly, one potentially preferred embodiment ofthe invention 18 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:46PM FROM-A J PARK Vo~n +64 4 4T23358 T-186 P-023/099 F-266 WO 02/28311 PCT/US0102304 0 may provide preparation of the sementhrough centrifugation. The semen may be diluted to provide, for example, 100 x 106 spermatozoa/ml in prefrably a commercial skim milk extender (EZ-Mixin CST@, Animal Reproduction Systems, Chino, CA). The sperm suspension may be protected from light and maintained for preferably 6 hours at room (20 25c) temperatre to simulate the potential time neededto sort the spermatozoa, if so desired.

The sperm suspension may then be centrifuged through a 90:45% Perooll (Sigma Chemical Co., St. Louis, MO, USA) discontinuous density gradient withthe goal ofreconcentrating the i cells and to potentially select ahighly motile fraction of spermatozoa- The 90% Percoll may be diluted at a ratio of 1:1 with HEPES-buffered Tyrode's medium (Grandahl et aL, o 10 1996) to make a 45% solution In a preferably 15-mt centrifuge tube, preferably 1 mL of o 45% Percoll may be carefully layered on top of preferably I mL of 90% Percoll. Preferably I mL ofthe spern suspension (100 x 10" sperm/mL inEZ-Miin, CST) maybe layered ontop ofthe Percoll layers, and the tube may be centritfged at 800 x g for a preferred period of 12 minutes. Afte centrifugation, the supematantmay be completelyremoved andthe pelletmay be resuspended in preferably 600gl HEPES-buffered Tyrode's Medium. The sperm concentration may be determined, in accordance with one embodiment, using a Densimeter (534B MOD-1, Animal Reproduction Systems, Chino, CA) and the potential volume to deliver 5 million spermatozoa (potentially of a preferred ~1000l) may be calculated and prepared for inseminationL In accordance with a broad aspect of the present invention, one embodiment thereof may provide for the positioning ofthe catheter near the UTJ ofthe female species ofmammal whereby the catheter is inserted under a surface in the vicinity of the uterotubaljunction. At least a portion of the sample may be extruded or deposited in the vicinity of the UTJ under the surface, One embodiment ofthepresent invention may provide for the collecting of spermcells from the male species of mammal, establishing an artificial insemination sample utilizing at least some of the sperm cells collected, and placing the artificial insemination sample in a catheter. A determination of a time when the female is appropriately fertile may be detemined, as described above. The optical element (12) andthe catheter (2)may be inserted 19 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:46PM FROM-A J PARK Vo~n +64 4 4723358 T-168 P-024/099 F-266 SWO 02/28311 PCT/US01/02304 Nvaginally and guided through the vagina, as depicted in Fig=e 2. The UTJ may be optically Slocated with optical element (12).

The catheter may be inserted under a surface (34) in the vicinity of the UTJ as depicted in Figure 3. At least a portion of said artificial insemination sample may be extruded I 5 under the surface within the vicinity of the UTJ, thus creating a "blister" with the sample n enshrouded between layers. Deposition of at least a portion of the artificial insemination Ssample may be provided Inder the surface (34) in the vicinity of the UTJ. The surface (34) N may comprise the endometrium or other portion of the uterus or the uterine lining. In o accordance with one preferred embodiment,the catheter may be inserted schthat aparticular 10 portion of the uterus is not pierced, and in accordance with one embodiment, such that a vasoularized portion of the uterus is not pierced. A vascularized portion of the uterus that may not be pierced, according to one particular embodiment, may include, for example, the mesometrium or the myometrium portion of the uterus or other vascularized portions.

Thereafter, the optical element and catheter may be withdrawn from the female and fertilization of an egg ofthe female may occur afterwhich production ofan offspring mammal may be expected from the fertilized egg.

An embodiment ofthe presentinventionmay also provide apotentially corresponding inseminationcatheter having a guide element, or inpreferred embodiments avideoeadoscope or a canula, a reservoir element responsive to the guide element, an extrusion element, or in preferred embodiments a syringetowhich saidreservoir element is responsive, and acellular piercing tip (36) positioned in front of the reservoir element The catheter may further provide a pierce depth control element, such as an adjustment element or a stop on the piercing tip (36) positioned in the vicinity of the tip. One embodiment of an insemination catheter of the present invention is depicted in Figure 3.

As previously described and in accordance with the insemination system ofextrusion and deposition under a surface in the vicinity of the UTJ described above, a number of steps of producing a mammal may be performed. A low number of sperm may be placed in the catheter and inpreferred embodiments, preferably numbers selected from: less than about COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:47PM FROM-A J PARK iton +64 4 4723358 T-168 P.025/099 F-266 WO 02/28311 PCT/US01/02304 0 (Nten million sperm, less than about five million sperm, less than about two million sperm, less Sthan about one million sperm, less than about five hundred thousand spenn, and less than about one hundred thousand sperm. Furthermore, the fertilization of an egg may be perfo=ned in accordance with the preferred embodiments ofthe invention wherein success levels success levels of fertilizationmaybe statistically comparable to a conventional uterine Sbody artificial insemination process. Statistically comparable success levels may be defined as previously mentioned.

Furthermore, the sperm cells may be collected from a male species of mammal, in It- I alternative embodiments ofthe invention, comprising bovids, equids, or swine. Inaccordance o 10 with altemative embodiments of the invention, sperm cells may be selected from collected cells for those cells that may be more likely to achieve insemination, as previously described.

Accordingly, an insemination containment element may be provided, in accordance with embodiments of the invention, preferably comprising a cellular base surface, and in particular embodiments a uterine lining or, in accordance with prefered embodiments, a nonvascularizd portion ofthe uterus, such as the mesometrium orthemyometrium, a cellular cover surface adjacent to the cellular base surface, and in particular embodiments, the endometrium oruterinelining, a substantial enshroudedvolumebetween the cellularbase and he cover surface, and inpreferred embodiments located inthe vicinity ofthe UTJ,and sperm cells from the male of the species. Preferred embodiments may also utilize low numbers of sperm relative to natural insemination, located within the volume and a sperm emission element adjacent the volume through which sperm may pass. The sperm may be collected, selected, of an inseminate volume, perhaps even of an epididymis origin, or of any other limitation previously discussed. The sperm emission element may comprise a breach in the endometrium surface of the uterus, as depicted in Figure 3, or may simply occur by diffusion or the like.

Additionally, other potentially independent procedures may be incorporated into the present invention and may still be considered as within the scope of the present invention.

Such procedures may include sorting the sperm cells by a sex characteristic, thereby 21 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:47PM FROM-A J PARK gton +64 4 4723358 T-168 P-026/099 F-266 SWO 02/28311 PCT/IS01/02304 0 (C establishing asex-sorted artifioial inseminationsample, and in preferredembodiments having Sa low number of sperm compared to a natural insemination dosage for said mammal, may include establishing a low dose sex-selected artificial insemination sample. Furtheomore, preserving or freezing, and the subsequent thawing of, sperm sells may be accomplished in particular embodiments, particularly inregardto various mammals such as equid, bovid and swine. Deposition ofthe inseminaion sample, may be processed or aspirated in any way, may be deposited with the crypts or folds of the UTJ, and may provide some type of preservation of the sperm for subsequent insemination. Establishment of an insemination specimen or 0 insemination sample atahysteroscopic compaible volume and iizing compatible mediamay o 10 further provide for allowing cooling of the specimen or sample at room temperature.

Centrifgationmay preferablybe perfrmedtrough aPercol gradientfor about five minutes at about 200g and for about ten minutes at about 800g. In accordance with a preferred embodiment, concentrating themore motile spermmay be limitedto concentatingto less than about twice the starting concentration. In particular, the broad and narrow concepts embodied in the present invention should be construed as applying to other species of mammal, including equids, bovids and swine. Finally the present invention, directed in part to the producing of an offspring mammal, may further be considered to disclose an embodiment of an animal produced utilizing a process as described in any of the foregoing method claims.

Sorting, in accordance with embodiments of the present invention, may particularly provide for collecting sperm cells fromamale of a species ofmammal, sorting the sperm cells according to a sex-specific characteristic, establishing a sorted, sex-specific artificial insemination sample, placing the sorted, sex-specific artificial insemination sample in a catheter; among the various other aspects of the invention disclosed herein that might be incorporated in method of producing a mammaL As can be easily understood from the foregoing, the basic concepts of the present invention maybe embodied in a variety of ways. It involves both insemination techniques as well as apparans to accomplish appropriate insomination. In this application, the insemination techniques are disclosed as part of the results shown to be achieved by the various devices 22 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:47PM FROM-A J PARK Vton 64 4 4723358 T-160 P.02T/099 F-266 WO 02/28311 PCT/US01/02304 0 ~N described and as steps which are inherentto utilization. They are simply the natural result of utilizing the devices as intended and described. In addition, while some devices are disclosed, it should be understood that these not only accomplish certainmethods but also canbe varied Sin a number of ways. Importantly, as to all of the foregoing, all of these facets should be understood to be encompassed by this disclosure.

The discussion included in this applicationis intended to serve as a basic description.

The reader should be aware that the specific discussion may not explicitly describe all C( embodimentspossible; many alternatives areimplicit. It also may not fully explainthe generic nature ofthe invention and may not explicitly showbow each feature or element can actually o 10 be representative of a broader function or of a great variety of alternative or equivalent elements. Again, these are implicitly included in this disclosure. Where the invention is described in device-oriented terminology, each element of the device implicitly performs a function. Apparatus claims may not only be included for the devices described, but also method or process claims may be included to address the functions the invention and each element performs. Neither the description nor the terminology is intendedto limit the scope of the claims which will be included in a full patent application.

It should also be understood that avariety of changes may be made without departing fromthe essence ofthe invention- Such changes are also implicitly includedinthe description.

They still fall within the scope of this invention.

Further, each ofthe various elements ofthe invention and claims may also be achieved in a variety of manners This disclosure should be understood to encompass each such variation, be it a variation of an embodiment of any apparatus embodiment, a method of process embodiment, or even merely a variation of any element of these. Particularly, it should be understoodthat as the disclosure relates to elements of the invention, the words for each elementmay be expressedby equivalent apparatus terms or method te rm s even if only the function or result is the same. Such equivalent, broader, or even more generic terms should be considered to be encompassed in the description of each element or action. Such terms can be substitutedwhere desiredto make explicit the implicitly broad coverage to which 23 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:48PM FROM-A J PARK Vaon +64 4 4723358 T-168 P-028/099 F-266 WO 0228311 PCT/US01/02304 WO OZ/28311 0 S this invention is entitled.

As but one example, it should be understood that all actions may be expressed as a means for taking that action or as an element which causes that action. Similarly, each physical element disclosed should be understood to encompass a disclosure of the action which that physical element facilitates. Regarding this last aspect, as but one example, the disclosure of an "extruder" should be understood to encompass disclosure of the act of "extruding" whether explicitly discussed or not and, conversely, were there only Sdisclosure of the act of "extruding", such a disclosure should be understood to encompass o disclosure of an "xtruder" and even a "means for extruding". Such changes and alternative o 10 terms are to be understood to be explicitly included in the description.

Any acts oflaw, statutes, regulations, or rules mentioned inthis applicationfor patent, or patents, publications, or other references mentioned in this application for patent, are hereby incorporated by reference. In addition, as to each term used it should be understood that unless its utilization in this application is inconsistent with such interpretation, common dictionary definitions should be understood as incorporated for each term and all definitions, alternative terms, and synonyms such as contained in the Random House Webster's Unabridged Dictionary, second edition are hereby incorporated by reference. However, as to each of the above, to the extent that such information or statements incorporated by reference might be considered inconsistent with thepatenting ofthis/these invention(s), such statements are expressly not to be considered as made by the applicant(s).

Thus, the applicant(s) should be understood to have support to claim at least: i) each ofthe insemination devices asherein disclosed and described, ii) threlatedmethods disclosed and described, iii) similar, equivalent, and even implicitvariations of each ofthese devices and methods, iv) those alternative designs which accomplish each of the functions shown as are disclosed and described, v) those alternative designs and methods which accomplish each of the functions shown as are implicit to accomplish that which is disclosed and described, vi) each feature, component, and step shown as separate and independent inventions, vii) the applications enhanced by the various systems or components disclosed, viii) the resulting 24 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:48PM FROM-A J PARK Vton +64 4 4723358 T-168 P-029/099 F-266 WOO2/28311 PCT/US01/02304 0 Sproducts produced by such systems or components, and ix) methods and apparatuses substantially as described hereinbefore and with reference to any of the accompanying examples, and x)the various combinations and pennutations ofeachofthe elements disclosed.

Further, if or when used, the use of the transitional phrase "comprising" is used to maintain the "open-end" claims herein, according to traditional claim interpretation. Thus, unless the context requires otherwise, it should be understood that the term "comprise" or variations such as "comprises" or"comprising", areintendedto imply the inclusion ofastated element or step or group of elements or steps but not the exclusion of any other element or Sstep or group of elements or steps. Such tens should be interpreted in their most expansive form so as to afford the applicant the broadest coverage legally permissible.

It should also be noted tat the term"at least one" as used inthe following description and claims is not intended nor used in this disclosure to menanthat other clais or descriptions not incorporating the "at least one" language cannot further include one or more like elements. More specifically, the language "at least one" is not intended nor used to change "open-ended" claims, inherently including devices or methods having additional elements or steps apart from those claimed, into "clo sed-ended" claims wherein devices ormetods having additional elements would not be covered by such claims. Accordingly, if or when used, the use ofthe transitional phrase "comprising" is usedto maintainthe "open-end" claims herein, according to traditional claim interpretation.

I. PATENT DOCUMENTS DOCUMENT NO. DATE NAME/COUNTR.Y CLASS SUBCLASS FLNG DAT_ 5,135,759 08/04/92 Johnson 424 561 04/26/91 6,071,689 06/06/00 Scidol et at 435 2 01129/98 601238,294 05/10/00 Morris et al. 05/O00 WO 98/34094 06/o0398 NZ WO 99/33956 08/07/99 US WO 99138883 05/08/99 US WO 99/42810 26108199 jUS WO 00/06193 10/02/00 US -1 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:48PM FROM-A J PARKgtn+4 4735T-6 P.0/9 F26 oton +64 4 4T23358 T-168 P-030/099 F-266 WO0/81 PCTJJJSGI/02304 IL OTHER DOCUMYIENTS (Jacludig Author, rTWO, Dale, Perfine~ut Pages, Bu.) Brhi, V, and Aen. WMt, Videcendoscopic Emrination of the Mare's Uterus: Findings inwornsl Fatie Mares", Equine Veternary Journal, Vol. 24 (1992% pp. 274-278 Buchmnan, B.It, vcal 4 Insamiot of Mares wit Low Numbers of~ither'Unsexed or Sexed Spetntozoa", S Talgeology, Vol. 53, pp 1333-1344, (2000) lflCaslick. FPA, "the Vulva and the Vulve-vagia Orifice and its Relation to Genital Health of &ce Thoroughbred Mare, Cornell Veterinarian. VoL 27,1931, -pp. 178-187 o Unsorted Semen", 'Theriogenclogy, Vol. 47, pp. 267, (AbstruAt), (1997) Pugor NY-lnical Experience with Flow Cytometiic Saeanrion of 11uma X- and Y- Cbnnnasome Bearing o Sperr", Theriogeziolozy, Vol. 52,1pp. 1435-1440 (1999) Ci GrandaM, C, vt al, 'in Vitro production ot'Rpine Emnbryos", Biology of Reproduction, Monograph Scrims 1 pp.

maximm efficiency", 'lberiosenulogy, Vol. 52, (1999), pp. 132S3-1341 Ziobnson, "Sex Pseseletico in Swine. Altered Sex Ratios in Offspring Following Surgical Insemnination of FMw Sorted X- e=4 Bearing Sperm"', Reproduction in pomestic Animals, Vol. 26, pp. 309-314 (1991) Johnson, et "Sex Prcction. in Rabbits. Live Birts trorn X and Y Sperm Separated by DNA a=d Cell Sortiag7 Biology of' Reproduction, Vol. 41, pp. 199-203 (1989) Manning S et al., "Development of Hysteroscopic, Insemination of the Uterine Tube in the Marc", Proceedings of the MnAprl Meeting of the Society forTheriogeaclogy. 1998, pp. 84-85preovulatorytiiarce', Journalof Reproduction ald FertlityVol 118, pp. 95-100 (2800>) Thberiogenology, Vol 44 619"27 (1995) Piocett OW, et OL, "Managemeat of the meare for niaximrnun reproductive Widency" Bulletin No. 6 Colorado State University. Ft Collins CO. (1989) Scheak, IL, "CryopreservaliOfl of flow-soxted bovine spcrwantozoC", Theriogenology, Vol. 52, 1375-1391 (1999) Schmnid ILL., et al, "Fertilization with Sexed Equine Spermatooa Ulsing IlrAcytolplasmic Sperm TIection and OviducWa Insemination ,7th Internationial Symposium On Equne Rgproduction, pp. 139 (Abstract) (1998) Sd],G. real,"tiCdIJernation of Ileifers with Cooled, Unfrozen Sexed Semen Theriogenclogy, Vol -49 pp- '365 (Abstract) (199 8) Seidel, G.E. Ir, etal., Iosegminndnfld of Heillow with Sexed Speraim Sheiogenology, VoL 52, pp. 1407-1421 (1999) 26 COMS ID No: SBMI-6947858 Received by IP Australia: Time (I-tm) 15:17 Date 2007-04-11 11-04-2007 04:4QPM FROM-A J1 PARKgtn+447238T18 P0 /O F-6 Von +64 4 4T23358 T-168 P-031/099 F-266 PCT/USOI/02304 VIO 02/22311 Squires. L, "laxy Bmbiyonie Loss "in Equine Diagsic ibtrsonography. 1' t E& w 157-I63 Eds Ltenen McY,,inn.William and Wilihns, BaltIMOrs MaxYland (1998) Squires ct "Cooled and ERozon stsahon sezne" Bulletin No,.9, Colorado Stateflnivesity, Ft. Collns, VqzuW!, L, etal., "AJ. in Swine; New Strategy for Deep Insemination with Low Number of Spermaoza Using a Non-surgical MethodolOeY, 14' Intmsaiional Congress an Animal Reproducdarn, VoL 2, Stocclom, Aily, 2000, p.

289.

Vazquez, et at,5 "osurgicil Utero-tuba Insemkofaioli in the Mare", Proceedings Ainual Meeting of the Society ibr Thcriogenfloly, 1998, pp. 82-43, Vazquez ct al,, "suceasildl Low-Dose Insemination by a Fiberoptie 13ndoscope Technique in tte Sow, Proceediinga Annual Conlference of the Intmrnutonal Embryo Transfer Society, Netherlands, Thedoganology, VoL 53, January, 2000, pp. 201.

Vaquas, L, mt at, seDevelopment of a onSlrgical Deep TVr= Uterie Insemination Tecniqite", IV intcrnalil Conference on Boar Samoa PreservaiVon, Maryland. August, 199.9, p 35 and photo Of display board.

Vmnquer at 4l. "Development of aLon-surgical Peep Yntin Uterine hactuinsfn Technique, Boar Semen Preservation IV, Wtl ternational Contferc= on, Boar Semen preservation, Maryland, pp. 262-263- Vqe', ct sl.,u' ypoosiotic Swelling Test as predctr of th embrae Integrt jnBoarSpennatoxo" 3oar Semen presevation IV, IVda Intentonal Confeyrnce on Boar Semen Prsevation, Maryland, pp. 263- Johnson, t al, "Sex Pregelection in swing: Flow Cytometic Sorting of K- and Y- ChromosoME Bearing Spelmnto Produce offTring", Bor Semeon Preservation IV, 2000, pp. 107-114.

Rath, bo.,e a "Low Dose isemination Tchnique in tire PNg, Boar Semen Preservtion TV, 2000, pp.11-19.

Lindse. et al., "Hlysteroscopic Insemination of Mares with Nonfrozu Low-dose Unstated or Sex-sorted.

Spermatczat', cuerently impublisbed. pp. 1-15.

21 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11

Claims (196)

11-04-2007 04:49PM FROM-A J PARK ffton +64 4 4723358 T-168 P-032/099 F-266 SWO 02/28311 PCTUS01/02304 0 0 The claims defining the invention are as follows: 1. A method of producing a mammal comprising the steps of: a. collecting sperm cells from a male of a species of mammal; 5 b. selecting desirable sperm for insemination; c. determining a hysteroscopic isemination compatible volume for artificial insemination; o' d. establishing an insemination specimen at said hysteroscopic insemination o compatible volume utilizing said desirable sperm for insemiation and utilizing hysteroscopic compatible media in said insemination specimen; e. temporally substantially retaining a character of said sperm cells in said insemination specimen; determining an estrous time for a female of a species of said mammal; g. altering a property of said insemination specimen to establish an artificial insemination sample at about said estrous time; h. placing said artificial insemination sample in a catheter; i. determining a time when a female of said species of said mammal is appropriately fertile; j. vaginally inserting an optical element into said female of said species of said mammal; k. vaginally inserting said catheter into a female of said species of said mammal; 1, guiding said optical element and said catheter through said vagina of said female of said species of said mammal; m. optically locating a uterotubal junction within said female of said species of said mammal; n. positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal; o. extruding at least a portion of said artificial insemination sample from said COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:49PM FROM-A J PARK Vton +64 4 4723358 T-168 P-033/099 F-266 WO 02/2831 PCT/US01/02304 catheter under a surface in the vicinity of said uterotubal junction within said female of said species of said mammal; p. depositing at least a portion of said artificial insemination sample in the vicinity of said uterotubal junction within said female of said species of said mammal; q. withdrawing said optical element and said catheter from said female of said species of said mammal; r. fertilizing an egg of said female of said species of said mammal; and s- producing an offspring mammal from said fertilized egg. 2. A method of producing a mammal as described in claim 1 wherein said step of fertilizing an egg of said female of said species of said mammal comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process. 3. A method of producing a mammal as described in claim 2 wherein said step of placing said artificial insemination sample in a catheter comprises the step of placing a low number of sperm in said catheter. 4. A method of producing a mammal as described in claim 3 wherein said step of placing a low number of sperm in said catheter comprises the step of placing a number of sperm selected from a group consisting of: less than about ten million sperm, less than about five million sperm, less than about two million sperm, less than about one million spem, less than about five hundred thousand sperm, and less than about one hundred thousand sperm. A method of producing a mammal as described in claim 4 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group 29 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:50PM FROM-A J PARK VOR 64 4 4723358 T-168 P.034/099 F-266 WO 028311 PCT/S01/02304 we W02128311 0 0 N consisting of: Sfertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination Sprocess with a statistical P value greater than or equal to at least about 0.05, ferilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination 0 process with a statistical P value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about 6. A method of producing a mammal as described in claim 4 wherein said step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process comprises the step of fertilizing an egg of said female of said species of said mammal with success levels selected from a group consisting of: at least about success rates, at least about 65% success rates, at least about 60% success rates, at least about 50% success rates, at least about 45% success rates, and at least about 90% of a conventional artificial insemination for that species. 7. A method of producing a mammal as described in claim 6 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of: fertilizing an egg of said female of said species of said mammal with success COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:50PM FROM-A J PARK Vlon 64 4 4723358 T-168 P-035/099 F-266 l WO 02/28311 PCT/US0102304 0 levels statistically comparable to a conventional uterine body artificial insemination Sprocess with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination 0 process with a statistical 3 value greater than or equal to at least about o 10 fertilizing an egg of said female of said species of said mammal with success o levels statistically comparable to a conventional uterine body artificial insemination process with both a satistical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about 8. A method ofproducing a mammal as described in claim 6 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine. 9. A method of producing a mammal as described in claim 8 wherein said step of positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal comprises the step of inserting said catheter under a surface in the vicinity of said uterotubal junction within said female of said species of said mammal, wherein said step of extending at least a portion of said artificial insemination sample from said catheter under said surface in the vicinity of said uterotubal junction within said female of said species of said mammal comprises the step of extruding at least a portion of said artificial insemination sample from said catheter under said surface in the vicinity of said uterotubal junction within said female of said species of said mammal, and wherein said step of depositing at least a portion of said aspirated insemination sample in the vicinity of said terotubal junction within said female of said species of said mammal within 31 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:50PM FROM-A J PARK Vo~n 64 4 4723368 T-168 P-036/099 F-266 I PCTIUSO1IO23o4 WO 022811 PCT/US01/02304 0 ci said female of said species of said mammal comprises the step of depositing at least Sa portion of said artificial insemination sample under said surface in the vicinity of said uiterotubal junction within said female of said species of said mammal within said female of said species of said mammal. 10. A method of producing a mammal as described in claim 8 wherein said step of establishing an insemination specimen at said hysteroscopic insemination compatible volume utilizing said desirable sperm for insemination and utilizing Shysteroscopic compatible media in said insemination specimen comprises the step Sof establishing an artificial insemination sample utilizing fresh sperm. 11. A method ofproducing a mammal as described in claim 6 wherein said step of depositing at least a portion of said aspirated insemination sample in the vicinity of said uterotubal junction within said female of said species of said mammal comprises the step of depositing at least a portion of said aspirated insemination sample within crypts of said uterotubal junction within said female of said species of said mammal.

12. A method of producing a mammal as described in claim 6 wherein said step of establishing an insemination specimen at said hysteroscopic insemination compatible volume utilizing said desirable sperm for insemination and utilizing hysteroscopic compatible media in said insemination specimen comprises the step of diluting said spern cells.

13. A method of producing a mmnial as described in claim 12 wherein said step of diluting said sperm cells comprises the step of diluting said spera cells selected- from a group consisting of: to no more than a 2:1 ratio, to no more than a 5:1 ratio, to no more than a 10:1 ratio, diluting with a skim milk medium, diluting with a TALP medium, diluting with a HEPES-buffered Tyrode's medium, and diluting with an Androhep medium. 32 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:51PM FROM-A J PARK ffton +64 4 4723358 T-168 P-0371/099 F-266 I1 PCTIUS01/02304 [WO 02/28311 0 S14. A method ofproducing a mammal as described in claim 13 wherein said step of establishing an insemination specimen at said hysteroscopic insemination compatible volume utilizing said desirable sperm for insemination and utilizing hysteroscopic compatible media in said insemination specimen fuTtber comprises the step of cooling said insemination specimen at about room temperature. A method of producing a mammal as described in claim 6 wherein said step of selecting desirable sperm for insemination comprises the step of concentrating the (N more motile sperm collected from said male species of said mammal. 0 S16. A method of producing a mammal as described in claim 15 wherein said step of concentrating the more motile sperm collected from said male species of said mammal comprises the step of cetrifuging said sperm through a Percoll gradient.

17. A method of producing a mammal as described in claim 16 wherein said step of centrifuging said sperm through a Percoll gradient comprises the step of utilizing a 90:45% Percoll gradient

18. A method of producing a mammal as described in claim 17 wherein said step of centrifuging said sperm through a Percoll gradient further comprises the steps of: a. centrifuging said sperm through said Percoll gradient for about five minutes at about 200g; and b. centrifuging said sperm through said Percoll gradient for about ten minutes atabout 80 g.

19. A method of producing a mammal as described in claim 17 wherein said step of concentrating the more motile sperm collected from said male species of said mammal comprises the step of limiting said concentration of said spean to less than about twice the starting concentration.

20. A method of producing a mammal as described in claim 6 wherein said step of 33 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:51PM FROM-A J PARK C~n +r64 4 4723358 T-168 P-038/099 F-266 0 0 Oc I< PCTUS01/02304 WO 02128311 establishing comprises the step of establishing an insemination specimen at a volume selected from a group consisting of: between about 30 and 150 ul, less than about 500 ul, about 230 u and about 100 ul.

21. A method of producing a mammal as described in claim 6 wherein said step of determining an estrous time for a female of a species of said manmmal comprises the step of inducing estrus in said female of said species of said mammal.

22. A method of producing a mammal as described in claim 21 wherein said step of inducing estrus in said female of said species of said mammal comprises the step of synchronously inducing estrus in a plurality of females of said species of said mammal. 23_ A method of producing a mammal as described in claim 21 or 22 wherein said step of inducing estrus in said female of said species of said mammal comprises the steps of: a. subjecting said female of said species of said mammal to daily treatments of progestagen for about ten days; and b. subjecting said female of said species of said mammal to at least one treatments of cloprostenol about the tenth days of accomplishing said step of subjecting said female of said species of said mammal to daily treatments of progestagen for about ten days.

24. A method of producing a mammal as described in claim 6 and further comprising the step of initiating said step of altering a property of said insemination specimen to establish an artificial insemination sample at about said estrous time at about the time determined for said female of said species of said mammal to be appropriately fertile.

25. A method of producing a mammal as described in claim 24 wherein said step of initiating said step of altering a property of said insemination specimen to establish 34 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:51PM FROM-A J PARK Vo~n +64 4 4T23358 T-168 P-039/099 F-266 WO 021283T1 PCTIS01/02304 0 CA an artificial insemination sample at about said estrous time at about the time Sdetermined for said female of said species of said mammal to be appropriately fertile comprises the step of initiating said step of altering a property of said insemination specimen within a time selected from a group consisting of; within about twenty-four hours of said time determined for said female of said species of said mammal to be appropriately fertile, within about twenty-four hours of said time determined for said female of said species of said mammal to be appropriately fertile, within about twelve hours of said time determined for said female of said 0 species of said mammal to be appropriately fertile, within about eight hours of said o 10 time determined for said female of said species of said mammal to be appropriately Sfertile, within about three hours of said time determined for said female of said species of said mammal to be appropriately fertile, and within about one hour of said time determined for said female of said species of said mammal to be appropriately fertile.

26. A method of producing a mammal as described in claim 6 wherein said step of guiding said optical element and said catheter through said vagina of said female of said species of said mammal comprises the steps of: a. guiding said optical element and said catheter through a cervix of said female of said species of said mammal; b. guiding said optical element and said catheter through a lumen of said female of said species of said mammal; and c. guiding said optical element and said catheter through a uterine horn of said female of said species of said mammal.

27. A method of producing a mammal as described in claim 26 wherein said step of guiding said optical element and said catheter through a cervix of said female of said species of said mammal comprises the step of manually guiding said optical element and said catheter through said cervix of said female of said species of said mammal, and wherein said step of guiding said optical element and said catheter through a lumen of said female of said species of said mammal comprises the step COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:52PM FROM-A J PARK Vton +64 4 4723358 T-168 P-040/099 F-266 WO 02/28311 PCT/US01/02304 of manually guiding said optical element and said catheter through said lumen of said female of said species of said mammal.

28. A method of producing a mammal as described in claim 26 and further comprising the step of inserting an illumination element in the vicinity of said optical element

29. A method of producing a mammal as described in claim 28 and further comprising the step of strobing said illumination element. A method of producing a mammal as described in claim 29 wherein said step of guiding said optical element and said catheter through a uterine born of said female of said species of said mammal comprises the step of optically guiding said optical element and said catheter through a uterine horn of said female of said species of said mammal.

31. A method of producing a mammal as described in claim 6 and further comprising the step of determing a dominant follical within said female of said species of said mammal.

32. A method of producing a mammal as described in claim 6 and further comprising the step of observing a preovulatory follicle of at least about 35 mm in size.

33. A method of producing a mammal as described in claim 6 wherein said step of positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal comprises the steps of a_ positioning said catheter within about three to five cm of said uterotubal junction; and then b. touching said catheter to said uterotubal junction.

34. A method of producing a mammal as described in claim 6 and further comprising the step of introducing filtered air with said uterus of said female of said species of 36 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:52PM FROM-A J PARK Kton 64 4 4723358 T-168 P.041/099 F-266 I WO 02/28311 PCT/US01/02304 0 Ssaid mammal. c| A method of producing a mammal as described in claim 34 and farther comprising the step of evacuating air from said uterus of said female of said species of said mammal.

36. A method ofproducing a mammal as described in claim 6 and further comprising the step of aspirating at least a portion of said artificial insemination sample 0 extruded from said catheter to create an aspirated insemination sample. o

37. A method ofproducing a mammal comprising the steps of: a. collecting sperm cells from a male of a species of mammal; b. establishing an artificial insemination sample having a low number of sperm compared to a natural insemination dosage for said mammal; c. placing said artificial insemination sample in a catheter, d. determining a time when a female of said species of said mammal is appropriately fertile; e. vaginally inserting an optical element into said female of said species of said mammal; f. vaginally inserting said catheter into a female of said species of said mammal; g. guiding said optical element and said catheter through said vagina of said female of said species of said mammal; k optically locating a uterotubal junction within said female of said species of said mammal; i. positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal; j. extruding at least a portion of said artificial insemination sample from said catheter; k. depositing a low number of sperm in the vicinity of said uterotubal junction within said female of said species of said mammal; 37 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:52PM FROM-A J PARK Vton 64 4 4723358 T-168 P-042/099 F-266 WO 02/28311 PCT/JS01/02304 0 1. withdrawing said optical element and said catheter from said female of said Sspecies of said mammal; fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process; and n- producing an offspring mammal from said fetilized egg.

38. A method of producing a mammal as described in claim 37 wherein said step of 0 placing a low number of sperm in said catheter comprises the step of placing a o number of sperm selected from a group consisting of: less than about ten million 0 10 sperm, less than about five million sperm, less than about two million sperm, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm

39. A method of producing a mammal as described in claim 38 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical p value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination 38 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:53PM FROM-A J PARK ffton +64 4 4723358 T-168 P-043/099 F-266 IWO 2/28311 PCT/USO102304 O (N process with both a statistical P value greater than or equal to at least about 0.05 Sand a statistical 3 value greater than or equal to at least about A method of producing a mammal as described in claim 38 wherein said step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process comprises the step of fertilizing an egg of said female of said species of said mammal with success levels selected from a group consisting of: at least about S 75% success rats, at least about 65% success rates, at least about 60% success o rates, at least about 50% success rates, at least about 45% success rates, and at least about 90% of a conventional artificial insemination for that species.

41. A method of producing a mammal as described in claim 40 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of. fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0,05, fertilizing an egg of said female of said species of said m mmaal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical (3 value greater than or equal to at least about 39 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:53PM FROM-A J PARK Co~n +64 4 4723358 T-168 P-044/099 F-266 WO 02/28311 PCTIUS01/02304

42. A method of producing a mammal as described in claim 40 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

43. A method of producing a mammal as described in claim 42 wherein said step of positioning said catheter in the vicinity of said uteratubal junction within said female of said species of said mammal comprises the step of inserting said catheter under a surface in the vicinity of said uterotubal junction within said female of said species of said mammal, wherein said step of extruding at least a portion of said artificial insemination sample from said catheter under said surface in the vicinity of said uterotubal junction within said female of said species of said mammal comprises the step of extruding at least a portion of said artificial insemination sample from said catheter under said surface in the vicinity of said uterotubal junction within said female of said species of said mammal, and wherein said step of depositing at least a portion of said aspirated insemination sample in the vicinity of said uterotabal junction within said female of said species of said mammal within said female of said species of said mammal comprises the step of depositing at least a portion of said artificial insemination sample under said surface in the vicinity of said uterotubal junction within said female of said species of said mammal within said female of said species of said mammal.

44. A method of producing a mammal as described in claim 42 wherein said step of establishing an insemination specimen at said hysteroscopic insemination compatible volume utilizing said desirable sperm for insemination and utilizing hysteroscopic compatible media in said insemination specimen comprises the step of establishing an artificial insemination sample utilizing fresh sperm. A method of producing a mammal as described in claim 40 and further comprising the step of selecting, from the sperm cells collected, those more likely to achieve insemination. COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:53PM FROM-A J PARK ffton +64 4 4723358 T-168 P.045/099 F-266 WO 0228311PCT/US01/02304 wo 02/25311 0 CN 46- A method of producing a mammal as described in claim 45 wherein said step of selecting desirable sperm for insemination comprises the step of concentrating the more motile sperm collected from said male species of said mammal.

47. A method of producing a mammal as described in claim 46 wherein said step of concentrating the more motile sperm collected from said male species of said mammal comprises the step of centrifuging said sperm through a Percoll gradient

48. A method of producing a mammal as described in claim 47 wherein said step of o centrifuging said sperm through a Percoll gradient comprises the step of utilizing a S90:45% Percoll gradient.

49. A method of producing a-mammal as described in claim 46 wherein said step of concentrating the more motile sperm collected from said male species of said mammal comprises the step of limiting said concentration of said sperm to less than about twice the starting concentration. A method of producing a mammal as described in claim 40 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

51. A method of producing a mammal as described in claim 42 wherein said step of establishing comprises the step of establishing an insemination specimen at a volume selected from a group consisting of: between about 30 and 150 ul, less than about 500 il, about 230 ul, and about 100 ul.

52. A method of producing a mammal as described in claim 40 wherein said step of collecting sperm cells from a male of a species ofmammal comprises the step of collecting epididymal sperm cells fom a male of a species of mammal. 41 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:54PM FROM-A J PARK Vton 64 4 4723358 T-168 P-046/099 F-266 SWO 02/28311 PCT/US1/02304 0 CN 53. A method of producing a mammal as described in claim 42 wherein said step of establishing an artificial insemination sample having a low number of sperm compared to a natural insemination dosage for said mammal comprises the step of Sestablishing an artificial insemination sample utilizing hysteroscopic compatible media.

54. A method of producing a mammal as described in claim 53 wherein said step of establishing an artificial insemination sample utilizing hysteroscopic compatible media comprises the step of utilizing a slim milk medium. 0 o 55. A method of producing a mammal as described in claim 53 wherein said step of establishing an artificial insemination sample utilizing hysteroscopic compatible media comprises the step of utilizing a medium selected from a group consisting of a TALP medium, a HEPES-buffered Tyrode's medium, and an androhep medium.

56. A method of producing a mammal as described in claim 53 wherein said step of establishing an artificial insemination sample utilizing hysteroscopic compatible media comprises the step of utilizing a media having a catheter coordinated viscosity.

57. A method of producing a mammal as described in claim 40 and farther comprising the step of aspirating at least a portion of said artificial insemination sample extruded from said catheter to create an aspirated insemination sample.

58. A method of producing a mammal as described in claim 40,97, 134, 147, 155, 180, 194, 212, 225 and further comprising the step of sorting said sperm cells by a sex characteristic, and wherein said step of establishing an artificial insemination sample having a low number of sperm compared to a natural insemination dosage for said mammal comprises the step of establishing a sex-selected artificial insemination sample having a low number of sperm compared to a natural insemination dosage for said mammal. 42 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:54PM FROM-A J PARK Vton 64 4 4723358 T-168 P.04T/099 F-266 SWO 02/28311 PCT/US01/02304 0 N 59. A method of producing a mammal as described in claim 58 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

60. A method of producing a mammal as described in claim 58 and further comprising the step of freezing said sperm cells.

61. A method of producing a mammal as described in claim 60 wherein said step of F collecting sperm cells from amale of a species of mammal comprises the step of o collecting sperm cells from a male of a species of a mammal selected from a group consisting of bovids, equids, or swine.

62. A method of producing a mammal as described in claim 40, 97, 134, 147, 155, 180, 194, 212, 225 and further comprising the step of freezing said sperm cells.

63. A method of producing a mammal as described in claim 62 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

64. A method ofproducing a mammal as described in claim 40, 294,235, 225, 212, 204, 194, 180, 155, 147, 134 wherein said step of depositing at least a portion of said aspirated insemination sample in the vicinity of said uterotabal junction within said female of said species of said mammal comprises the step of depositing at least a portion of said aspirated insemination sample within crypts of said uterotubal junction within said female of said species of said mammal. A method of producing a mammal as descrbed in claim 40, 294,235, 225, 212, 204, 194, 180, 155, 147, 134 and further comprising the step of facilitating the adherance of said sperm to a surface in the vicinity of said uterotubal junction 43 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:54PM FROM-A J PARK ffton +64 4 4723358 T-168 P-048/099 F-266 I 02/2831 PCTUS01/02304 WO 02/2831 0 N within said female of said species of said mammal.

66. A method of producing a mammal as described in claim 65 wherein said step of Sfacilitating the adherance of said sperm to a surface in the vicinity of said uterotubal junction within said female of said species of said mammal comprises the step of creating a froth from at least a portion of said artificial insemination sample.

67. A method of producing a mammal as described in cdaim 40, 294,235, 225, 212, 204, 194, 180, 155, 147, 134 and further comprising the step of creating a froth o" from at least a portion of said artificial insemination sample.

68. A method of producing a mammal as described in claim 40,294,235,225, 212, 204, 194, 180, 155, 147, 134, 97 wherein said step of establishing an insemination specimen at said hysteroscopic insemination compatible volume utilizing said desirable sperm for insemination and utilizing hysteroscopic compatible media in said insemination specimen comprises the step of diluting said sperm cells.

69. A method of producing a mammal as described in claim 68 wherein said step of diluting said sperm cells comprises the step of diluting said sperm cells selected from a group consisting of: to no more than a 2:1 ratio, to no more than a 5:1 ratio, to no more than a 10:1 ratio, diluting with a skim milk medium, diluting with a TALP medium, diluting with a HEPES-buffered Tyrode's medium, and diluting with an Androhep medium.

70. A method of producing a mammal as described in claim 69 wherein said step of establishing an insemination specimen at said hysteroscopic insemination compatible volume utilizing said desirable sperm for insemination and utilizing hysteroscopic compatible media in said insemination specimen furthe comprises the step of cooling said insemination specimen at about room temperature.

71. A method of producing a mammal as described in claim 40, 294, 235, 225, 212, 44 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:54PM FROM-A J PARK gton +844 4 4723358 T-168 P-049/099 F-266 WO 0228311 PCT/USO1/02304 O 204, 194, 180, 155, 147, 134, 97 and further comprising the step of selecting, from the sperm cells collected, those more likely to achieve insemination.

72. A method of producing a mammal as described in claim 7lwhrein said step of selecting desirable sperm for insemination comprises the step of concentrating the more motile sperm collected from said male species of said mammal. I t

73. A method of producing a mammal as descnibed in claim 72 wherein said step of Sconcentrating the more motile sperm collected from said male species of said Smammal comprises he step of centrifuging said sperm through a Percoll gradient

74. A method of producing a mammal as described in claim 73 wherein said step of centrifuging said sperm through a Percoll gradient comprises the step of tiizing a 90:45% Percoll gradient. A method of producing a mammal as described in claim 72, 74 wherein said step of centrifuging said sperm through a Percoll gradient further comprises the steps of: a. centrifuging said sperm through said Percoll gradient for about five minutes at about 200g; and b. centrifuging said sperm through said Percoll gradient for about ten minutes at about 800g.

76. A method ofproducing a mammal as described in claim 72 wherein said step of concentrating the more motile sperm collected from said male species of said mammal comprises the step of limiting said concentration of said sperm to less than about twice the starting concentration.

77. A method of producing a mammal as described in claim 72 wherein said step of establishing comprises the step of establishing an insemination specimen at a volume selected fom a group consisting of: between about 30 and 150 ul, less than about 500 ul, about 230 ul, and about 100 ul. COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:55PM FROM-A J PARK ffton +64 4 4723358 T-168 P-050/099 F-266 0 0 ci WO 02/28311 PCT/US01/02304

78. A method of producing a mammal as described in claim 40,294,235,225,212, 204, 194, 180, 155, 147, 134, 97 wherein said step of determining an estrous time for a female of a species of said mammal comprises the step of inducing estrous in said female of said species of said mammal.

79. A method of producing a mammal as described in claim 78 wherein said step of inducing estrous in said female of said species of said mammal comprises the steps of: a. subjecting said female of said species of said mammal to daily treatments of progestagen for about ten days; and b. subjecting said female of said species of said mammal to at least one treatments of cloprostenol about the tenth days of accomplishing said step of subjecting said female of said species of said mammal to daily treatments of progestagen for about ten days. A method of producing a mammal as described in claim 40, 294, 235, 225, 212, 204, 194, 180, 155, 147, 134, 97 and further comprising the step of initiating said step of altering a property of said insemination specimen to establish an artificial insemination sample at about said estrous lime at about the time determined for said female of said species of said mammal to be appropriately fertile.

81. A method ofproducing a mammal as described in claim 80 wherein said step ofintiating said step of altering a property of said insemination specimen to establish an artificial insemination sample at about said estrous time at about the time determined for said female of said species of said mammal to be appropriately fertile comprises the step of initiating said step of altering aproperty of said insemination specimen within a time selected from a group consisting of: within about twenty-four hours of said time determined for said female of said species of said mammal to be appropriately fertile, within about twenty-four hours of said time determined for said female of said species of said mammal to be appropriately fertile, within about twelve hours of said time determined for said female of said 46 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:55PM FROM-A J PARK Cton +64 4 4723358 T-168 P-051/099 F-266 SWO 02/28311 pCT/US01/02304 0 species of said mammal to be appropriately fertile, within about eight hours of said Stime determined for said female of said species of said mammal to be appropriately fertile, within about three hours of said time determined for said female of said species of said mammal to be appropriately fertile, and within about one hour of said time determined for said female of said species of said mammal to be l appropriately fertile. In S82. A method of producing a mammal as described in claim 40, 294, 235, 225,212, S 204, 194, 180, 155, 147, 134, 97 wherein said step of guiding said optical element o and said catheter through said vagina of said female of said species of said mammal C 10 comprises the steps of: a. guiding said optical element and said catheter through a cervix of said female of said species of said mammal; b. guiding said optical element and said catheter through a lumen of said female of said species of said mammal; and c. guiding said optical element and said catheter through a uterine horn of said female of said species of said mammal.

83. A method of producing a mammal as described in claim 82 wherein said step of guiding said optical element and said catheter through a cervix of said female of said species of said mammal comprises the step of manually guiding said optical element and said catheter through said cervix of said female of said species of said mammal, and wherein said step of guiding said optical element and said catheter through a lumen of said female of said species of said mammal comprises the step of manually guiding said optical element and said catheter through said lumen of said female of said species of said mammal.

84. A method of producing a mammal as described in claim 82 and further comprising the step of inserting an illumination element in the vicinity of said optical element. A method of producing a mammal as described in claim 84 and further comprising 47 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:55PM FROM-A J PARK gton +64 4 4T23358 T-168 P.052/099 F-266 SWO 02/28311 PCTJS01/02304 0 the step of strobing said illumination element S86. A method of producing a mammal as described in claim 85 wherein said step of Sguiding said optical element and said catheter through a uterine horn of said female of said species of said mammal comprises the step of optically guiding said optical I 5 element and said catheter through a uterine horn of said female of said species of i/t saidmammal 0 C 87. A method of producing a mammal as described in claim 40,294,235,225,212, 0204,194,180,155,147, 13 4,97 and farther conmprising the step ofdetemning a Ci dominant follical within said female of said species of said mammal.

88. A method of producing a mammal as described in claim 40,294,235,225,212, 204, 194, 180, 155, 147, 134,97 and further comprising the step of observing a preovulatory folicle of at least about 35 tum in size.

89. A method of producing a mammal as described in claim 40,294,235,225,212, 204,194,180,155,147, 134, 97 wherein said step of positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal comprises the steps of: a. positioning said catheter within about three to five cm of said uterotubal junction; and then b. touching said catheter to said uterotubal junction.

90. A method of producing a mammal as described in claim 40,294,235, 225,212, 204, 194, 180, 155, 147, 134, 97 and fmrther comprising the step of introducing filtered air with said uterus of said female of said species of said mammal.

91. A method ofproducing a mammal as described in claim 90 and ft u rther comprising the step of evacuating air from said uterus of said female of said species of said mammunaL 48 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:56PM FROM-A J PARK ton +64 4 47Z3358 T-168 P.053/099 F-266 WO 02/28311 PCT/US01/02304 0 C 92, A method of producing a mammal as described in claim 40,294,235,225,212, O-i 204, 194, 180,155,147, 134,97 and futher comprising the step of aspirating at least a portion of said artificial isemination sample extruded from said catheter to create an aspirated insemination sample.

93. A method of producing a mammal comprising the steps of: Sa. collecting sperm cells from a male of a species of mammal; b. establishing an artificial insemination sample utilizing at least some of said c sperm cells; c placing said artificial insemination sample in a catheter, 1 0 d. determining a time when afemale of said species of said mammal is appropriately fertile; e. vaginally inserting an optical element into said female of said species of said mammal; f. vaginally inserting said catheter into a female of said species of said mammal; g. guiding said optical element and said catheter through said vagina of said female of said species of said mammal; h. optically locating a uterotubal junction within said female of said species of said mammal; i. inserting said catheter under a surface in the vicinity of said uterotubal junction within said female of said species of said mammal; j. extrding at least a portion of said artificial insemination sample from said catheter under said surface in the vicinity of said uterotubal junction within said female of said species of said mammal; k. depositing at least a portion of said artificial insemination sample under said surface in the vicinity of said uterotubal junction within said female of said species of said mammal within said female of said species of said mammal; 1. withdrawing said optical element and said catheter from said female of said species of said Mammal; m. fertilizing an egg of said female of said species of said mammal; and 49 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:56PM FROM-A J PARK gton +64 4 4723358 T-168 P-04/099 F-266 WO 02/28311 PCT/US01/02304 0 C, n. producing an offspring mammal from said fertilized egg.

94. A method of producing a mammal as described in claim 93 wherein said step of placing said artificial insemination sample in a catheter comprises the step of placing a low number of sperm in said catheter.

95. A method of producing a mammal as described in claim 94 wherein said step of o placing a low number of sperm in said catheter comprises the step of placing a number of sperm selected from a group consisting of: less than about ten million o sperm, less than about five million sperm, less than about two million sperm, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand spenm.

96. A method of producing a mammal as described in claim 95 wherein said step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process comprises the step of fertilizing an egg of said female of said species of said mammal with success levels selected from a group consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:56PM FROM-A J PARK ffton +64 4 4723358 T-168 P-055/099 F-266 WO 02/28311 PCT/US01/02304 0 C and a statistical p value greater than or equal to at least about

97. A method of producing a mammal as described in claim 95 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels selected from a group consisting of: at least about 75% success rates, at least about 65% success rates, at o least about 60% success rates, at least about 50% success rates, at least about Nsuccess rates, and at least about 90% of a conventional artificial insemination for lr- 0 that species. 0

98. A method of producing a mammal as described in claim 97 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical 3 value greater than or equal to at least about ferilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical 3 value greater than or equal to at least about 51 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:57PM FROM-A J PARK V~n +64 4 4723358 T-168 P-056/099 F-266 WO 02/28311 PCTIUS01/02304 0 C 99- A method of producing a mammal as described in claim 97 wherein said step of Scollecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

100. A method of producing a mammal as described in claim 93 and further comprising Sthe step of selecting, from the sperm cells collected, those more likely to achieve Sinsemination. o

101. A method of producing a mammal as described in claim 100 wherein said step of 0 selecting desirable sperm for insemination comprises the step of concentrating the more motile sperm collected from said male species of said mammal.

102. A method of producing a mammal as described in claim 101 wherein said step of concentrating the more motile sperm collected from said male species of said mammal comprises the step of centrifuging said sperm through a Percoll gradient.

103. A method of producing a mammal as described in claim 102 wherein said step of centrifuging said sperm through a Percoll gradient comprises the step of utilizing a 90:45% Percoll gradient

104. A method of producing a mammal as described in claim 101 wherein said step of concentrating the more motile sperm collected from said male species of said mammal comprises the step of limiting said concentration of said sperm to less than about twice the starting concentration.

105. A method of producing a mammal as described in claim 93 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting spern cells from a male of a species of a mammal selected from a group consisting of- bovids, equids, or swine. 52 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:57PM FROM-A J PARK ffton +64 4 4T23358 T-168 P-05/099 F-266 WO 02/28311 PCT/US01/02304

106. A method of producing a mammal as described in claim 93 wherein said step of establishing comprises the step of establishing an insemination specimen at a volume selected from a group consisting of. between about 30 and 150 ul, less than about 500 ul, about 230 l, and about 100 ul.

107. A method of producing a mammal as described in claim 93 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting epididymal sperm cells from a male of a species ofmammal.

108. A method of producing a mammal as described in claim 93 wherein said step of inserting said catheter under a surface in the vicinity of said uterotubal junction within said female of said species of said mammal comprises the step of piercing an endomitiumn surface of auterus within said female of said species of said mammal.

109. A method of producing a mammal as described in claim 108 wherein said step of inserting said catheter under a surface in the vicinity of said uterotubal junction within said female of said species of said mammal further comprises the step of avoiding piercing a vascularized portion of said uterus of said female of said species of said mammal.

110. A method of producing a mammal as described in claim 93 wherein said step of piercing an endomitrium surface within a uterus of said female of said species of said mammal comprises the step of tlizing said catheter.

111. Amethod of producing a mammal as describedin claim 110 and further cmpriing the step of controlling the depth to which said catether pierces said endomitrium surface of said utems within said female of said species of said mammal.

112. A method of producing a mammal as described in claim 111 wherein said step of controlling the depth to which said catether pierces said endomitrium surface of said uterus within said female of said species of said mammal comprises the step of 53 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-200T1 04:57PM FROM-A J PARK Vton 64 4 4723358 T-168 P-058/099 F-266 WO 02/28311 PCT/US01/02304 Sutilizing said catheter.

113. An insemination containment element comprising: a. a cellular base surface; b. a cellular cover surface adjacent to said cellular base surface; lt) 5 c. a substantially enshrouded volume between said cellular base surface and tfl said cellular cover surface; Sd. spenn cells from amale of a species of mammal located within said c substantially enshrouded volume; and O e. a sperm emission element adjacent said substantially enshrouded volume Ci 10 and through which said sperm cells can pass.

114. An insemination containment element as described in claim 113 wherein said sperm cells from a male of a species of maotal located within said substantially enshrouded volume comprise a low number of sperm cells as compared to athe number of spemn cells typically found in a natural insemination-

115. An insemination containment element as described in claim 114 wherein said low number of sperm cells as compared to a the number of sperm cells typically found in a natural insemination comprises a number of sperm cells selected from a group consisting of: less than about ten million sperm, less than about five million sperm, less than about two million sperm, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm.

116. An insemination containment element as described in claim 115 wherein said sperm cells comprise sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

117. An insemination containment element as described in claim 113 wherin said sperm cells from a male of a species of mammal located within said substantially 54 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:58PM FROM-A J PARK ton 64 4 4723358 T-168 P-059/099 F-266 WO 02/28311 PCT/US01/0230 enshrouded volume comprise sperm cells selected from sperm cells collected which Sare more likely to achieve insemination. 7

118. An insemination containment element as described in claim 117 wherein said sperm cells selected from sperm cells collected which are more likely to achieve insemination comprise sperm cells selected to be the more motile sperm collected in from said male species of said mammal. O 1 S119. An insemination containment element as described in claim 113 wherein said sperm o cells from a male of a species of mammal located within said substantially f^ enshrouded volume comprise fresh sperm cells.

120. An insemination containmejt element as described in claim 113 wherein said sperm cells from a male of a species of mammal located within said substantially enshrouded volume has an inseminate volume and wherein said inseminate volume comprises a volume selected from a group consisting of: between about 30 and 150 u less than about 500 u, about 230 ul, and about 100 ul.

121. An insemination containment element as described in claim 113 wherein said sperm cells from a male of a species ofmammal located within said substantially enshrouded volume comprise epididymal sperm cells from a male of a species of mammal.

122. An insemination containment element as described in claim 115 wherein said sperm cells from a male of a species of manmmal located within said substantially enshrouded volume comprise sex-selected sperm cells from a male of a species of mammal.

123. An insemination containment element as described in claim 122 wherein said sex- selected sperm cells from a male of a species of mammal comprise sperm cells from a male of a species of a mamnal selected from a group consisting of: bovids, COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:58PM FROM-A J PARK Vton +64 4 4T23358 T-168 P-060/099 F-266 SWO 02/28311 PCTUS01/02304 0 0 equids, or swine.

124. An insemination containment element as described in claim 115 wherein said Ssubstantially enshrouded volume between said cellular base surface and said cellular cover surfaceis located in the vicinity of a uterotubal junction within a female of said species of said mammal. S125. An insemination containment element as described in claim 124 wherein said CN cellular cover surface adjacent to said cellular base surface comprises an o endomitrium surface of a uterus within said female of said species of said mammal. 0

126- An insemination containment element as described in claim 125 wherein said cellular base surfacecomprises a vascularized portion of said uterus of said female of said species of said nammal.

127. An insemination containment element as described in claim 115 wherein said sperm emission element adjacent said substantially enshrouded volume and through which said sperm cells can pass comprises a breach in said endomitrium surface of said uterus within said female of said species of said mammal.

128. An insemination catheter comprising: a. a guide element; b. a reservoir element responsive to said guide element; c. an extrusion element to which said reservoir element is responsive; and d. a cellular piercing tip positioned in front of said reservoir element.

129. An insemination catheter as described in claim 128 and further comprising a pierce depth control element positioned in the vicinity of said cellular piercing tip.

130. A method of producing a mammal comprising the steps of a. collecting sperm cells from a male of a species of mammal; 56 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:58PM FROM-A J PARK Vton 64 4 4T23358 T-168 P.061/099 F-266 WO 02/28311 pCTIUS01102304 b. selecting, from the sperm cells collected, those more likely to achieve inseminatiorn c. establishing an artificial insemination sample using said desirable sperm for insemination; d placing said artificial insemination sample in a catheter; e. determining a time when a female of said species of said mammal is appropriately fertile; f vaginally inserting an optical element into said female of said species of said mammal; g. vaginally inserting said catheter into a female of said species of said mammal; h. guiding said optical element and said catheter through said vagina of said female of said species of said mammal; i. optically locating a uterotaal junction within said female of said species of said mammal; j. positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal; k. extruding at least a portion of said artificial insemination sample from said catheter under a surface in the vicinity of said uterotubal junction within said female of said species of said mammal; 1. depositing at least a portion of said artificial insemination sample in the vicinity of said uterotubal junction within said female of said species of said mammal within said female of said species of said mammal; m. withdrawing said optical element and said catheter from said female of said species of said mammal; n. fertilizing an egg of said female of said species of said mammal; and o. producing an offspring mammal from said fertilized egg.

131. A method of producing a mammal as described in claim 130 wherein said step of placing said artificial insemination sample in a catheter comprises the step of placing a low number of sperm in said catheter. 57 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:58PM FROM-A J PARK ffton 64 4 4723358 T-168 P-062/099 F-266 WO 02/28311 PCT/US01/02304

132. A method of producing a mammal as described in claim 131 wherein said step of placing a low number of sperm in said catheter comprises the step of placing a number of sperm selected from a group consisting of: less than about ten million sperm, less than about five million sperm, less than about two million sperm, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm.

133. A method of producing a mammal as described in claim 132 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical p value greater than or equal to at least about

134. A method of producing a mammal as described in claim 132 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels selected from a group 58 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:59PM FROM-A J PARK Vton +64 4 4723358 T-168 P.063/099 F-266 WO OV28311 PCT/US01/02304 consisting of: at least about 75% success rates, at least about 65% success rates, at least about 60% success rates, at least about 50% success rates, at least about success rates, and at least about 90% of a conventional artificial insemination for that species.

135. A method of producing a mammal as described in claim 134 wherein said step of fertilizing comprises the step offctilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterie body artificial insemination process with success levels determined from a group consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insmination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical 0 value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical value greater than or equal to at least about

136. A method of producing amammal as described in claim 134 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from amale of a species ofa mammal selected from a group consisting of: bovids, equids, or swine.

137. A method of producing a mammal as described in claim 134 wherein said step of 59 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:59PM FROM-A J PARK irtan 64 4 423358 T-168 P-064/099 F-266 PCT/USOl/02304 0 0 WO 02/28311 selecting desirable sperm for insemination comprises the step of concentrating the more motile sperm collected from said male species of said mammal.

138. A method of producing amammal as described in claim 137 wherein said step of concentrating the more motile sperm collected from said male species of said mammal comprises the step of centrifuging said sperm through a Percoll gradient.

139. A method of producing a mammal as described in claim 138 wherein said step of centrifging said sperm through a Percoll gradient comprises the step of utilizing a 90:45% Percoll gradient.

140. A method of producing a mammal as described in claim 137, 139 wherein said step of centrifuging said sperm through a Percoll gradient further comprises thn steps of: a. cetrifugig said spenn through said Percoll gradient for about five minutes at about 200g; and b. centrifuging said sperm through said Percoll gradient for about ten minutes at about 800g.

141. A method of producing amammal as described in claim 137 wherein said step of concentrating the more motile sperm collected from said male species of said mammal comprises the step of limiting said concentration of said sperm to less than about twice the starting concentration.

142. A method of producing a mammal as described in claim 137 wherein said step of establishing comprises the step of establishing an insemination specimen at a volume selected from a group consisting of: between about 30 and 150 ul, less than about 500 ul, about 230 l, and about 100 ul.

143. A method of producing a mammal comprising the steps of: a. collecting sperm cells from a male of a species of mammal; COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 04:59PM FROM-A J PARK VOR +64 4 4T23358 T-168 P-065/099 F-266 pCT/US01102304 0" "WO 02/28311 C /2304 0 C b. establishing an artificial insemination sample utilizing fresh sperm; i .0 placing said artificial insemination sample in a catheter; d. determining a time when a female of said species of said mammal is appropriately fertile; e. vaginally inserting an optical element into said female of said species of said Smammal; tl f. vaginally inserting id catheter into a female of said species of said 0 mammal; g. guiding said optical element and said catheter through said vagina of said o 10 female of said species of said mammal; Sh. optically locating a terotubal junction within said female of said species of said mammal; i. positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal; j. extruding at least a portion of said artificial insemination sample from said catheter; k. depositing at least a portion of said artificial insemination sample in the vicinity of said uterotubal junction within said female of said species of said mammal; 1. withdrawing said optical element and said catheter from said female of said species of said mammal; m. fertilizing an egg of said female of said species of said mammal; and n. producing an offspring mammal from said fertilized egg.

144. A method of producing a mammal as described in claim 143 wherein said step of placing said artificial insemination sample in a catheter comprises the step of placing a low number of sperm in said catheter.

145. A method of producing a mammal as described in claim 144 wherein said step of placing a low number of sperm in said catheter comprises the step of placing a number of sperm selected from a group consisting of: less than about ten million 61 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:00PM FROM-A J PARK ffton 64 4 4T23358 T-168 P.066/099 F-266 SWO 028311 PCT/USO1/02304 Ssperm, less than about five million sperm, less than about two million sperm, less Sthan about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm.

146. A method of producing a mammal as described in claim 145 wherein said step of t 5 fertilizing comprises the step of fertilizng an egg of said female of said species of t said mammal with success levels statistically comparable to a conventional uterine Sbody artificial insemination process with success levels determined from a group consisting of: o fertilizing an egg of said female of said species of said mammal with success C 10 levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fetilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical value greater than or equal to at least about

147. A method of producing a mammal as described in claim 145 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels selected from a group consisting of: at least about 75% success rates, at least about 65% success rates, at least about 60% success rates, at least about 50% success rates, at least about success rates, and at least about 90% of a conventional artificial insemination for 62 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:00PM FROM-A J PARK Vton +64 4 4723358 T-168 P.067/099 F-266 WO 02/28311 PCT/US01/02304 that species.

148. A method of producing amammal as described in claim 147 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical 3 value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about

149. A method of producing a mammal as described in claim 147 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a ale of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

150. A method of producing a mammal as described in claim 143 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting epididymal sperm cells from a male of a species of mammal. 63 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:OPM FROM-A J PARK ffton +64 4 4723358 T-168 P-068/099 F-266 WO 02/2311 PCT/US01/02304 C

151. A method of producing a mammal comprising the steps of: a collecting sperm cells from a male of a species of mammal; b. establishing an artificial insemination sample utilizing hysteroscopic compatible media; c. placing said artificial insemination sample in a catheter; VC) d. determining a time when a female of said species of said mammal is Sappropriately fertile; e. vaginally inserting an optical element into said female of said species of said O C mammal; o 10 f. vaginally inserting said catheter into a female of said species of said o mammal; g. guiding said optical element and said catheter through said vagina of said female of said species of said mammal; h. optically locating a uterotubal junctionwithin said female of said species of said mammal; i. positioning said catheter in the vicinity of said uterotubal junctio within said female of said species of said mammal; j. extruding at least a portion of said artificial insemination sample from said catheter, k. depositing at least a portion of said artificial insemination sample in the vicinity of said uterotubal junction within said female of said species of said mammal; 1. withdrawing said optical element and said catheter from said female of said species of said mammal; m. fertilizing an egg of said female of said species of said mammal; and n. producing an offspring mammal from said fertilized egg.

152. A method of producing a mammal as described in claim 151 wherein said step of placing said artificial insemination sample in a catheter comprises the step of placing a low number of spem in said catheter. 64 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:01PM FRO-A J PARK Vton +64 4 4723358 T-168 P-069/099 F-266 WO 02/28311 CT/US01102304

153. A method of producing a mamnal as described in claim 152 wherein said step of placing a low number of sperm in said catheter comprises the step of placing a number of sperm selected from a group consisting of: less than about ten million sperm, less than about five million sperm, less than about two million sperm, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm.

154. A method of producing a mammal as described in claim 153 wherein said step of Sfertilizing comprises the step of ferilizing an egg of said female of said species of Ssaid mammal with success levels statistically comparable to a conventional uterine 0 10 body artificial insemination process with success levels determined from a group consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical 3 value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insaeination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical p value greater than or equal to at least about

155. A method of producing a mammal as described in claim 153 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels selected from a group COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:01PM FROM-A J PARK ffton +64 4 4723358 T-168 P-070/099 F-266 WO 02/28311 PCT/USO1/0230 0 (N consisting of: at least about 75% success rates, at least about 65% success rates, at least about 60% success rates, at least about 50% success rates, at least about success rates, and at least about 90% of a conventional artificial insemination for _that species.

156. A method of producing a mammal as described in claim 155 wherein said step of IC fertilizing comprises the step of fertilizing an egg of said female of said species of Ssaid mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group Sconsisting of: 0 10 fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equalto at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about

157. A method of producing a mammal as described in claim 155 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

158. A method of producing a mammal as described in claim 151 wherein said step of 66 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-200T 05:01PM FROMt-A J PARK ton +64 4 4723358 T-168 P-071/099 F-266 WO 0128311 PCTUS01/02304 0 N establishing an artificial insemination sample utilizing hysteroscopic compatible Smedia comprises the step of utilizing a skim milk medium. I< S159. A method ofprodcing a mammal as describedin claim 151 wherein said step of establishing an artificial insemination sample utilizing hysteroscopic compatible I 5 media comprises the step of utilizing a medium selected from a group consisting of i a TALP medium, a HEPES-buffered Tyrode's medium, and an androhep medium. N 160. A method of producing a mammal as described in claim 151 wherein said step of o establishing an artificial insemination sample utilizing hysteroscopic compatible 0 media comprises the step of utilizing a media having a catheter coordinated viscosity.

161. A method of producing a mammal as described in claim 160 wherein said step of utilizing a media having a catheter coordinated viscosity comprises the step of utilizing a media which creates an artificial insemination sample having a viscosity of greater than about that of the blood of said mammaL

162. A method of producing a mammal as described in claim 161 wherein said step of utilizing a media having a catheter coordinated viscosity further comprises the step ofutiliing a media which creates an artificial insemination sample having a viscosity of greater than about that of a saline solution.

163. A method of producing amammal as described in claim 161 wherein said step of utilizing a media having a catheter coordinated viscosity comprises the step of utilizing a media selected from a group consisting of: a media having a viscosity of greater than about 10Ocp, a media having a viscosity of greater than about 300cp, a media having a viscosity of greater than about 1000cp, a media having a viscosity of greater than about 3000cp, and a media having a viscosity of greater than about 6000cp, each at about the mammal's average body temperature- 67 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:02PM FROM-A J PARK Vton 64 4 4723358 T-168 P-072/099 F-266 WO 02/28311 PCT/US01/02304

164. A hysteroscopic insemination sample comprising: a. a reservoir element; b. a catheter system to which said reservoir element is responsive; G. a plurality of sperm cells contained within said reservoir element; and d. hysterscopic compatible media contained within said reservoir element and to which said sperm cells are responsive.

165. An insemination containment element as described in claim 164 wherein said plurality of sperm cells contained within said reservoir element comprise a low number of sperm cells as compared to a the number of sperm cells typically found in a natural insemination-

166- An insemination containment element as described in claim 165 wherein said low number of sperm cells as compared to a the number of sperm cells typically found in a natural insemination comprises a number of sperm cells selected from a group consisting of: less than about ten million sperm, less than about five million sperm, less than about two million spern, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm.

167. An insemination containment element as described in claim 166 wherein said sperm cells comprise sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

168. An insemination containment element as described in claim 166 wherein said plurality of sperm cells contained within said reservoir element comprise sex- selected sperm cells from a male of a species of mammal.

169. An insemination containment element as described in claim 168 wherein said sex- selected sperm cells from a male of a species of mammal comprise sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, 68 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:02PM FROM-A J PARK ffton 64 4 4723358 T-168 P-073/099 F-266 WO 02/2831) PCT/US01102304 equids, or swine.

170. A hysteroscopic inseminafio sample as described in claim 164 wherein said hysterscopic compatible media contained within said reservoir element and to which said sperm cells are responsive comprises a skim milk medium.

171. A hysteroscopic insemination sample as described in claim 164 wherein said hysterscopic compatible media contained within said reservoir element and to which said sperm cells are responsive comprises a medium selected from a group consisting ofa TALP medium, a HEPES-buffered Tyrode's medium, and an androhep medium.

172- A hysteroscopic insemination sample as described in claim 164 wherein said hysterscopic compatible media contained within said reservoir element and to which said sperm cells are responsive comprises a catheter coordinated viscosity media.

173. A hysteroscopic insemination sample as described in claim 172 wherein said catheter coordinated viscosity media comprises a media which creates an artificial insemination sample having a viscosity of greater than about that of the blood of said mammal.

174. A hysteroscopic insemination sample as described in claim 173 wherein said catheter coordinated viscosity media further comprises a media which creates an artificial insemination sample having a viscosity of greater than about that of a saline solution.

175. A hysteroscopic insemination sample as described in claim 172 wherein said catheter coordinated viscosity media further comprises amedia selected from a group consisting of:: a media having a viscosity of greater than about 100cp, a media having a viscosity of greater than about 300cp, a media having a viscosity of 69 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-20071 05:02PM FROM-A J PARK Cton +64 4 4723358 T-168 P-074/099 F-266 WO 02/28311 PCT/US01/02304 0 greater than about 1000cp, a media having a viscosity of greater than about 3000cp, and a media having a viscosity of greater than about 6000cp, each at about the mammal's average body temperature.

176. A method of producing a mammal comprising the steps of: a. collecting spemn cells from a ale of a species of mammal; b. determining a hysteroscopic insemination compatible volume for artificial insemination; Sc. establishing an artificial insemination sample at said hysteroscopic 0 insemination compatible volume; 0 10 &d placing said artificial insemination sample in a catheter; e. determining a time when a female of said species of said mammal is appropriately fertile; vaginally inserting an optical element into said female of said species of said mammal; g. vagially inserting said catheter into a female of said species of said mammal; Ih guiding said optical element and said catheter through said vagina of said female of said species of said mammal; i. optically locating a uterotubal junction within said female of said species of said mammal; j. positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal; k. extruding at least a portion of said artficial insemination sample from said catheter; 1. depositing at least a portion of said artificial insemination sample in the vicinity of said uterotbal junction within said female of said species of said mammal; m. withdrawing said optical element and said catheter from said female of said species of said mammal; n. fertilizing an egg of said female of said species of said mammal; and COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:03PM FROM-A J PARK ton +64 4 4723358 T-168 P.075/099 F-266 S02/28311 PCTIUS01/02304 0 o. producing an offspring mammal from said fertilized egg, 177, A method of producing a mammal as described in claim 176 wherein said step of placing said artificial insemination sample in a catheter comprises the step of placing a low number of sperm in said catheter.

178. A method of producing a mammal as described in claim 177 wherein said step of placing a low number of sperm in said catheter comprises the step of placing a number of sperm selected from a group consisting of: less than about ten million o spanm, less than about five million sperm, less than about two million sperm, less 0 than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm.

179. A method of producing a mammal as described in claim 178 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 71 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-20071 05:03PM FROM-A J PARK Vton +64 4 4723358 T-168 P.076/099 F-266 I WO 02/28311 PCT/US0l/02304 0 N and a statistical j3 value greater than or equal to at least about lg0 A method of producing a mammal as described in claim 178 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of I said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levls selected from a group o consisting of; at least about 75% success rates, at least about 65% success rates, at least about 60% success rates, at least about 50% success rates, at least about o success rates, and at least about 90% of a conventional artificial insemination for Sthat species.

181. A method of producing a mammal as described in claim 180 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

182. A method of producing a mammal as described in claim 180 wherein said step of establishing comprises the step of establishing an insemination specimen at a volume selected from a group consisting of between about 30 and 150 ul, less than about 500 ul, about 230 ul, and about 100 ul.

183. A hysteroscopic insemination sample comprising: a. a reservoir element; b. a catheter system to which said reservoir element is responsive; and c. a hysteroscopic compatible volume of sperm contained within said reservoir element

184. An insemination containment element as described in claim 183 wherein said hysteroscopic compatible volume of sperm contained within said reservoir element comprise a low number of speam cells as compared to a the number of sperm cells 72 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:03PM FROM-A J PARK ffton +c64 4 4723358 T-168 P- 077/099 F-266 WO 02/283 1 PCT/US01/02304 O ci typically found in a natural iasemination.

185. An insemination containment element as described in claim 184 wherein said low Snumber of sperm cells as compared to athe number of sperm cells typically found in a natural insemination comprises a number of sperm cells selected from a group consisting of: less than about ten million sperm, less than about five million spean, tf less than about two million sperm, less than about one million sprnn, less than Sabout five hundred thousand sperm, and less than about one hundred thousand C sperm. 0

186. An insemination containment element as described in claim 185 wherein said sperm cells comprise sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

187. A hysteroscopic insemination sample as described in claim 183 wherein said hysteroscopic compatible vohlme of sperm contained within said reservoir element comprises avolume selected from a group consisting of- between about 30 and 150- ul, less than about 500 ul, about 230 uL and about 100 ul.

188. An insemination containment element as described in claim 183,187 wherein said hysteroscopic compatible volume of sperm contained within said reservoir element comprise sex-selected sperm cells from a male of a species of mammal.

189. An inseminaion containment element as described in claim 188 wherein said sex- selected sperm cells from a male of a species of mammal comprise sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

190. A method of producing a mammal comprising the steps of: a. collecting sperm cells from a male of a species of mammal; b. temporally substantially retaining a character of said sperm cells in an 73 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:03PM FROM-A J PARK Vo~n 64 4 4T23358 T-168 P-078/099 F-266 WO 02/28311 PCT/US01/02304 0 insemination specimen; c. determining an estrous time for a female of a species of said mammal; d. altering a property of said insemination specimen to establish an artificial insemination sample at about said estros time; e. placing said artificial insemination sample in a catheter; l f. vaginally inserting an optical element into said female of said species of said mammal; Sg. vaginally inserting said catheter into a female of said species of said N' mammal; o- 10 b guiding said optical element and said catheter through said vagina of said fi female of said species of said mammal; i. optically locating a uterotubal junction within said female of said species of said mammal; j. positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal; k. extruding at least a portion of said artificial insemination sample from said catheter; 1, depositing at least aportion of said artificial insemination sample in the vicinity of said uterotabal junction within said female of said species of said mammal; m. withdrawing said optical element and said catheter from said female of said species of said mammal; n. fertilizing an egg of said female of said species of said mammal; and o. producing an offspring mammal from said fertilized egg.

191. A method of producing a mammal as described in claim 190 wherein said step of placing said artificial insemination sample in a catheter comprises the step of placing a low number of sperm in said catheter.

192. A method of producing a mammal as described in claim 191 wherein said step of placing a low number of sperm in said catheter comprises the step of placing a 74 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:04PM FROM-A J PARK ffton 64 4 473358 T-168 P-079/099 F-266 SWO 02128311 PCTIUSOI/02304 0 number of sperm selected from a group consisting of: les than about ten million sperm, less than about five million sperm, less than about two million sperm, less Sthan about one million sperm, less than about five hundred thousand sperm, and Sless than about one hundred thousand spermn IC 5 193. A method of producing a mammal as described in claim 192 wherein said step of I- fertilizing comprises the step of fertilizing an egg of said female of said species of Ssaid mammal with success levels statistically comparable to a conventional uterine C( body artificial insemination process with success levels determined from a group Sconsisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical p value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about

194. A method of producing a mammal as described in claim 192 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels selected from a group consisting of: at least about 75% success rates, at least about 65% success rates, at least about 60% success rates, at least about 50% success rates, at least about COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-200T1 05:4PM FROM-A J PARK ffton 64 4 4T23358 T-168 P-080/099 F-266 WO 02/28311 PCT/US01/02304 '0 succestess rates, and at least about 90% of a conventional artificial insemination for .that species. S195. A method of producing a mammal as described in claim 194 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting off fertilizing an egg of said female of said species of said mammal with success o levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical 3 value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about

196. A method of producing a mammal as described in claim 194 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

197. A method of producing a mammal as described in claim 190, 194, 196 and further comprising the step of initiating said step of altering a property of said insemination specimen to establish an artificial insemination sample at about said estrous time at 76 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-200T 05:04PM FROM-A J PARK Vo~n 64 4 4723358 T-168 P-081/099 F-266 WO 0/28311 PCT/USOl/02304 0 (N about the time determined for said female of said species of said mammal to be Sappropriately fertile. 1<

198. A method of producing a mammal as described in claim 197 wherein said step of initiating said step of altering a property of said insemination specimen to establish an artificial insemination sample at about said estrous time at about the time determined for said female of said species of said mammal to be appropriately fertile comprises the step of initiating said step of altering a property of said insemination specimen within a time selected from a group consisting of: within o about twenty-four hours of said time determined for said female of said species of said mammal to be appropriately fertile, within about twenty-four hours of said time determined for said female of said species of said mammal to be appropriately fertile, within about twelve hours of said time determined for said female of said species of said mammal to be appropriately fertile, within about eight hours of said time determined for said female of said species of said mammal to be appropriately fertile, within about three hours of said time determined for said female of said species of said mammal to be appropriately fertile, and within about one hour of said time determined for said female of said species of said mammal to be appropriately fertile.

199. A method of producing a mammal as described in claim 190, 194 wherein said step of temporally substantially retaining a character of said sperm cells in an insemination specimen comprises the step of freezing said sperm cells and wherein said step of altering a property of said insemination specimen to establish an artificial insemination sample at about said estrous time comprises the step of thawing said insemination specimen at about said estrous time.

200. A method of producing a mammal comprising the steps of: a. collecting sperm cells from a male of a species of mammal; b. sorting said sperm cells according to a sex-speciic characteristic; c. establishing a sorted, sex-specific artificial insemination sample; 77 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-200T1 05:05PM FROM-A J PARK Cton 64 4 4723358 T-168 P.082/099 F-266 WO 0218311 PCT/US01/02304 ad. placing said sorted, sex-specfic arificial insemination sample in a catheter; Se. determining a time when a female of said species of said mammal is <appropriately fertile; f. vaginally inserting an optical element into said female of said species of said mammal; g. vaginally inserting said catheter into a female of said species of said mammal; h. guiding said optical element and said catheter through said vagina of said 0. 'female of said species of said mammal; i. opticaily locating a uterotubal junction within said female of said species of said mammal; j. positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal; k. extruding at least a portion of said sorted, sex-specific artificial insemination sample from said catheter; 1. depositing at least a portion of said sorted, sex-specific artificial insemination sample in the vicinity of said uterotubal junction within said female of said species of said mammal; m. withdrawing said optical element and said catheter from said female of said species of said mammal; n. fertilizing an egg of said female of said species of said mammal; and o. producing an desired sex offspring mammal from said fertilized egg.

201. A method of producing a mammal as described in claim 200 wherein said step of placing said artificial insemination sampl in a catheter comprises the step of placing a low number of sperm in said catheter.

202. A method of producing a mammal as described in claim 201 wherein said step of placing a low number of sperm in said catheter comprises the step of placing a number of sperm selected from a group consisting of: less than about ten million sperm, less than about five million sperm, less than about two million spern,.less 78 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:05PM FROMA J PARK fftcn 64 4 4723358 T-168 P-083/099 F-266 WO 02/28311 PCTIUS01/02304 I than about one million sperm, less than about five hundred thousand sperm, and Sless than about one hundred thousand sperm.

203. A method of producing a mammal as described in claim 202 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of: Sfertilizing an egg of said female of said species of said mammal with success Slevels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical 3 value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about

204. A method of producing a mammal as described in claim 202 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels selected from a group consisting of at least about 75% success rates, at least about 65% success rates, at least about 60% success rates, at least about 50% success rates, at least about success rates, and at least about 90% of a conventional artificial insemination for that species. 79 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:05PM FROM-A J PARK Vton 64 4 4723358 T-168 P-084/099 F-266 SWO 02/28311 PCT/US01/02304 0 S205. A method of producing a mammal as described in claim 204 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group Sconsisting of: bovids, equids, or swine. 5 206. A method of producing amammal as described in claim 204 and further comprising the step of freezing said sperm cells.

207. A method of producing a mammal as described in claim 206 wherein said step of 0 collecting sperm cells from a male of a species of mammal comprises the step of Scollecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

208. A method of producing a mammal comprising the steps of: a. collecting epididymal sperm cells from a male of a species of mammal; b. establishing an artificial insemination sample; c. placing said artificial insemination sample in a catheter; d. determining a time when a female of said species of said mammal is appropriately fertile; e. vaginally inserting an optical element into said female of said species of said mammal; f. vaginally inserting said catheter into a female of said species of said mammal; g. guiding said optical element and said catheter through said vagina of said female of said species of said mammal; h. optically locating a uterotubal junction within said female of said species of said mammal; i. positioning said catheter in the vicinity of said uterotbal junction within said female of said species of said mammal; j. extruding at least a portion of said artificial insemination sample from said catheter; COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:06PM FROMA J PARK gton +64 4 4723358 T-169 P-085/099 F-266 WO 02/28311 PCTIUSO0/02304 0 Sk. depositing at least a portion of said artificial insemination sample in the Svicinity of said uterotubal junction within said female of said species of said mammal; L. withdrawing said optical element and said catheter from said female of said species of said mammal; m. fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process; and (N n. producing an offspring mammal from said fertilized egg. 0 010

209. A method of producing a mammal as described in claim 208 wherein said step of placing said artificial insemination sample in a catheter comprises the step of placing a low number of sperm in said catheter.

210. A method of producing a mammal as described in claim 209 wherein said step of placing a low number of sperm in said catheter comprises the step of placing a number of sperm selected from a group consisting of: less than about ten million sperm, less than about five million sperm, less than about two million sperm, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm

211. A method of producing a mammal as described in claim 210 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination 81 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-200T 05:06PM FROM-A J PARK MR 64 4 4723358 T-168 P-086/099 F-266 WO 02/28311 PCT/US01/02304 process using a 500 million equine sperm sample with a statistical P value greater than or equalto at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial inseination process with a statistical p value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about O 10 212. A method of producing a mammal as described in claim 210 wherein said step of fetilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels selected from a group consisting of: at least about 75% success rates, at least about 65% success rates, at least about 60% success rates, at least about 50% success rates, at least about success rates, and at least about 90% of a conventional artificial insemination for that species.

213. A method of producing a mammal as described in claim 212 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, 82 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:06PM FROM-A J PARK Vton 64 4 4723358 T-168 P-08T/099 F-266 SWO 02/2311PCT/USO/2304 S0 C fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical p value greater than or equal to at least about Sfertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination iprocess with both a statistical P value greater than or equal to at least about 0.05 and a statistical value greater than or equal to at least about C

214. A method of producing a mammal as described in claim 212 wherein said step of o collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

215. A hysteroscopic insemination sample comprising: a. a reservoir element; b. a catheter system to which said reservoir element is responsive; and c. a plurality of epididymal sperm cells contained within said reservoir element.

216. An insemination containment element as described in claim 215 wherein said plrality of epididymal sperm cells contained within said reservoir element comprise a low number of sperm cells as compared to a the number of sperm cells typically found in a natural insemination.

217. An insemination containment element as described in claim 216 wherein said low number of sperm cells as compared to a the number of sperm cells typically found in a natural insemination comprises a number of sperm cells selected from a group consisting of less than about ten million sperm, less than about five million sperm, less than about two million sperm, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm. 83 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:07PM FROM-A J PARK gton 64 4 4723358 T-168 P-088/099 F-266 WO 02/28311 PCT/US01/02304 0 CN 218. An insemination containment element as described in claim 217 wherein said sperm Scells comprise sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

219. An insemination containment element as described in claim 217 wherein said plurality of epididymal sperm cells contained within said reservoir element Scomprise sex-selected sperm cells from a male of a species of mammal

220. An insemination containment element as described in claim 219 wherein said sex- o selected sperm cells from a male of a species of mammal comprise sperm cells from Sa male of a species of a mmmal selected from a group consisting of bovids, equids, or swine.

221. A method of producing a mammal comprising the steps of: a. collecting sperm cells from a male of a species of mammal b. establishing an artificial insemination sample; c. placing said artificial insemination sample in a catheter; d, determining a time when a female of said species of said mammal is appropriately fertile; e. vaginally inserting an optical element into said female of said species of said mammal; f. vaginally inserting said catheter into a female of said species of said mammal; g. guiding said optical element and said catheter through said vagina of said female of said species of said mammal; h. optically locating a uterotubal junction within said female of said species of said mammal; i. positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal; j. extruding at least a portion of said artificial insemination sample from said catheter; 84 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-200T1 05:7PM FROM-A J PARK Vo~n +64 4 4723358 T-168 P-089/099 F-266 WO 02/28311 PCT/US01/02304 0 SIk. aspirating at least a portion of said artificial insemination sample extruded 1 from said catheter to create an aspirated insemination sample; 1. depositing at least a portion of said aspirated insemination sample in the vicinity of said terotubal junction within said female of said species of said mammal; m- withdrawing said optical element and said catheter from said female of said species of said mammal; n, fertilizing an egg of said female of said species of said mammal; and S'o. producing an offspring mammal from said fertilized egg. O 10 222. A method of producing a mammalas described in claim 221 wherein said step of placing said artificial insemination sample in a catheter comprises the step of placing a low number of sperm in said catheter. 223 A method of producing a mammal as described in claim 222 wherein said step of placing a low number of sperm in said catheter comprises the step of placing a number of sperm selected from a group consisting of less than about ten million sperm, less than about five million sperm, less than about two million sperm, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm.

224. A method of producing a mammal as described in claim 223 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:7PM FROM-A J PARK Vton 64 4 4723358 T-168 P-090/099 F-266 WO 02/28311 PC/US01/02304 0 process using a 500 million equine sperm sample with a statistical P value greater Sthan or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical 3 value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about O 10 225. A method of producing a mammal as described in claim 223 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels selected from a group consisting of; at least about 75% success rates, at least about 65% success rates, at least about 60% success rates, at least about 50% success rates, at least about success rates, and at least about 90% of a conventional artificial insemination for that species.

226. A method of producing a mammal as described in claim 225 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group consisting of. fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterne body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, 86 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:08PM FROM-A J PARK Vo~n +64 4 4723358 T-168 P-091/099 F-266 SWO 02/28311 PCT/USOI/02304 fertilizing an egg of said female of said species of said mammal with success Slevels statistically comparable to a conventional uterine body artificial insemination process with a statistical 3 value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statstical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about

227. A method of producing a mammal as described in claim 225 wherein said step of Scollecting sperm cells from a male of a species of mammal comprises the step of O 10 collecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

228. A hysteroscopic insemination element comprising: a. a uterus of a female species of a mammal; b. a plurality of sperm cells contained within said uterus; and c. an aspirated volume of media surrounding said sperm cells and to which said sperm cells are responsive.

229. An insemination containment element as described in claim 228 wherein said plurality of sperm cells contained within said uterus comprise a low number of sperm cells as compared to a the number of sperm cells typically found in a natural insemination.

230. An insemination containment element as described in claim 229 wherein said low number of sperm cells as compared to a the number of sperm cells typically found in a natural insemination comprises a number of sperm cells selected from a group consisting of: less than about ten million sperm, less than about five million sperm, less than about two million sperm, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand spermn 87 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:0PM FROM-A J PARK Vton +64 4 4723358 T-168 P-092/099 F-266 WO 02/28311 PCT/US01/02304 0 S231. An insemination containment element as described in claim 230 wherein said sperm cells comprise sperm cells from a male of a species of a mammal selected fom a group consisting of: bovids, equids, or swine. 232 An insemination containment element as described in claim 230 wherein said plurality of sperm cells contained within said uterus comprise sex-selected sperm cells from a male of a species of mammal.

233. An insemination containment element as described in claim 232 wherein said sex- selected sperm cells from a male of a species of mammal comprise sperm cells from O a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine.

234. A method of producing a mammal comprising the steps of a. collecting sperm cells from a male of a pig; b. establishing an artificial insemination sample having a low number of sperm compared to a natural insemination dosage for said pig; c. placing said artificial insemination sample in a catheter; d. determining a time when a female pig is appropriately fertile; e. vaginally inserting an optical element into said female pig; f. vaginally inserting said catheter into a female pig; g. guiding said optical element and said catheter through said vagina of said female pig; h. optically locating auterotubal junction within said female pig; i. positioning said catheter in the vicinity of said uterotubal junction within said female pig; j. extruding at least a portion of said artificial insemination sample from said catheter; k. depositing a low number of sperm in the vicinity of said uterotubal junction within said female pig; 1. withdrawing said optical element and said catheter from said female pig; 88 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-200TI 05:08PM FROM-A J PARK Vton 64 4 4T23358 T-168 P-093/099 F-266 WO 02/28311 PCT/US01/02304 0 fC m. fertilizing an egg of said female pig; and n. producing an offspring pig from said fertilized egg.

235. A method of producing a mammal as described in claim 234 wherein said step of fertilizing an egg of said female of said species of said mammal comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process. 0" 236. A method of producing a mammal as described in claim 235 and further comprising 0 the step of sorting said sperm cells by a sex characteristic, and wherein said step of establishing an artificial insemination sample having a low number of sperm compared to a natural insemination dosage for said mammal comprises the step of establishing a sex-selected artificial insemination sample having a low number of sperm compared to a natural insemination dosage for said mammal. 237 A method of producing a mammal as described in claim 236 and further comprising the step of freezing said sperm cells. 238 A method ofproducing a mamma l as described in claim 235 and further comprising the step of freezing said sperm cells. 239 A method of producing a mammal comprising the steps of: a. collecting sperm cells from a male of a species of mammal; b. freezing at least some of said sperm cells to create a frozen insemination sample; c. thawing at least some of said frozen insemination sample to create an artificial insemination sample; d. placing said artificial insemination sample in a catheter; e. determiing a time when a female of said species of said mammal is appropriately fertile; 89 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 0:0PM FROM-A J PARK Cton +64 4 4723358 T-168 P-094/099 F-266 WO 02/8311 PCT/US01/02304 0 f. vaginally inserting an optical element into said female of said species of said mammal; g. vaginally inserting said catheter into a female of said species of said mammal; h. guiding said optical element and said catheter through said vagina of said female of said species of said mammal; i. optically locating a uterotubal junotion within said female of said species of said mammal; j. positioning said catheter in the vicinity of said uterotubal junction within said female of said species of said mammal; k. extruding at least a portion of said artificial insemination sample from said catheter; 1. depositing at least a portion of said artficial insemination sample in the vicinity of said uterotubal junction within said female of said species of said mammal; m. withdrawing said optical element and said catheter from said female of said species of said mammal; n, fertilizing an egg of said female of said species of said mammal; and- o. producing an offspring mammal from said fertilized egg.

240. A method of producing a mammal as described in claim 239 wherein said step of fertilizing an egg of said female of said species of said mammal comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process.

241. A method of producing a mammal as described in claim 239, 240 wherein said step of placing said artificial insemination sample in a catheter comprises the step of placing a low number of sperm in said catheter.

242. A method of producing a mammal as described in claim 241 wherein said step of COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-200T 05:O9PM FROM-A J PARK ffton +64 4 4T23358 T-168 P-095/099 F-266 WO 02/28311 PCT/US01/02304 0 Splacing a low number of sperm in said catheter comprises the step of placing a Snumber of sperm selected from a group consisting of: less than about ten million sperm, less than about five million spenn, less than about two million sperm, less than about one million sperm, less than about five hundred thousand sperm, and less than about one hundred thousand sperm.

243. A method of producing a mammal as described in claim 242 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of o said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group o 10 consisting of: fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical 3 value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artiicial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about

244. A method of producing a mammal as described in claim 242 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels selected from a group consisting of: at least about 75% success rates, at least about 65% success rates, at 91 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-2007 05:0PM FROMA J PARK ffton 64 4 4723358 T-168 P.096/099 F-266 SWO 02/28311 PCT/US01/02304 0 least about 60% success rates, at least about 50% success rates, at least about Ssuccess rates, and at least about 90% of a conventional artificial insemination for that species.

245. A method of producing a mammal as described in claim 244 wherein said step of fertilizing comprises the step of fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with success levels determined from a group O consisting of: 0 fertilizing an egg of said female of said species of said mammal with success O 10 levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process using a 500 million equine sperm sample with a statistical P value greater than or equal to at least about 0.05, fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with a statistical P value greater than or equal to at least about fertilizing an egg of said female of said species of said mammal with success levels statistically comparable to a conventional uterine body artificial insemination process with both a statistical P value greater than or equal to at least about 0.05 and a statistical P value greater than or equal to at least about

246. A method of producing a mammal as described in claim 244 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of collecting sperm cells from a male of a species of a mammal selected from a group consisting ot bovids, equids, or swine.

247. A method of producing a mammal as described in claim 244 and further comprising the step of sorting said sperm cells by a sex characteristic, and wherein said step of 92 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11 11-04-200T1 05:10PM FRO-A J PARK ffton +64 4 4723358 T-168 P.0T/099 F-266 4WO 02/2311 PCT/US01/02304 O 0' establishing an artificial insemination sample having a low number of sperm compared to a naural insemination dosage for said mammal comprises the step of establishing a sex-selected artificial insemination sample having a low number of sperm compared to a natural insemination dosage for said mammal.

248. A method of producing a mammal as described in claim 247 wherein said step of collecting sperm cells from a male of a species of mammal comprises the step of Scollecting sperm cells from a male of a species of a mammal selected from a group consisting of: bovids, equids, or swine. O

249. An animal produced utilizing a process as described in any of the foregoing method claims. 93 COMS ID No: SBMI-06947858 Received by IP Australia: Time 15:17 Date 2007-04-11