CN1170511C - Compositions for freezing dog sperm, method of freezing the dog sperm utilizing the compositions and artificial insemination method employing the frozen dog sperm - Google Patents
Compositions for freezing dog sperm, method of freezing the dog sperm utilizing the compositions and artificial insemination method employing the frozen dog sperm Download PDFInfo
- Publication number
- CN1170511C CN1170511C CNB008080003A CN00808000A CN1170511C CN 1170511 C CN1170511 C CN 1170511C CN B008080003 A CNB008080003 A CN B008080003A CN 00808000 A CN00808000 A CN 00808000A CN 1170511 C CN1170511 C CN 1170511C
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- Prior art keywords
- sperm
- dog
- freezing
- compositions
- seminal fluid
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D19/00—Instruments or methods for reproduction or fertilisation
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D19/00—Instruments or methods for reproduction or fertilisation
- A61D19/02—Instruments or methods for reproduction or fertilisation for artificial insemination
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
- A61K35/52—Sperm; Prostate; Seminal fluid; Leydig cells of testes
Abstract
The invention discloses compositions for freezing dog sperm, a freezing method of the dog sperm utilizing the compositions and an artificial insemination method employing the frozen dog sperm are disclosed. The composition for freezing dog sperm contains 0.25 to 4.5 g of sodium citrate, 0.45 to 4.0 g of dextrose, 0.01 to 0.3 g of penicillin, 50,000 to 500,000 IU of streptomycin, 0.25 to 0.45 of catalase, 0.045 to 0.25 g of yolk, 1.5 to 15 ml of glycerol and 0.02 to 0.1 mu g of dog serum. The freezing method of the dog sperm includes the steps of (a) collecting male dog sperm by a massage method or electrical shock, (b) centrifuging the collected sperm to separate away the upper fluid and to fractionate only the lower sperm, (c) mixing the lower sperm and the composition in a volumetric ratio of 1:1 and primarily refrigerating the same, (d) further adding the composition to the mixture obtained in the step (c) to mix the sperm and the composition in a volumetric ratio of 1:2 and secondarily refrigerating the same, and (e) adding liquid nitrogen to the mixture obtained in the step (d) and freezing the same.
Description
Technical field
The present invention relates to be used for the compositions of frozen dog sperm, use the method for said composition frozen dog sperm and use this frozen dog sperm to carry out test-tube method.More specifically, the present invention relates to be used for the compositions of frozen dog sperm, wherein after the freezing of semen in the scheduled time with semen thawing, the quantity of survival sperm and shape thereof and survival ability remain unchanged in the seminal fluid, and the method for using the said composition frozen dog sperm, with test-tube method, it comprises refrigerated dog sperm is thawed in suitable place and time, and is inserted into female dog vagina.
Background technology
According to up-to-date industrial development, genetic engineering has become an important field of research.Genetic engineering must make good kind preserve and development, and human longevity is prolonged, and engineered research has exceeded the starting stage, has reached a quite high level.
For domestic animal and raising pets, the artificial insemination of using liquid seminal fluid can make several jennies inseminations by mixing with diluent from the seminal fluid that same buck is collected for the volume of the liquid of injecting to increase.Therefore,, can collect excellent or pure seminal fluid, and several jennies are fertilized simultaneously, can produce good kind thus and preserve pure kind by carrying out artificial insemination.
In order successfully to obtain artificial insemination with above-mentioned several advantages, developing proper diluent has become a major issue.Owing to begun the research of diluent before the many decades, developed the diluent that is used for pig, horse, cattle at present, and dropped into application.And the diluent that is used for dog sperm is studied immaturity still.
The fertilization of jenny need be used a kind of long-time preservation animal semen and not lose the method for the freshness of excellent seminal fluid, and this method can play important contribution effect to the development of domestic animal and house pet industry.
Freezing method is suitable for preserving for a long time sperm most.Yet, also specifically do not develop the freezing method of dog sperm.Although not fully aware of, the untapped reason that goes out the dog sperm freezing method may be to fail to develop fully to be used for freezing reagent with dog sperm sperm of special nature, and uses this compositions and methods.
Disclosure of the Invention
First purpose of the present invention is to be provided for the compositions of frozen dog sperm, can freezingly have the dog sperm of special sperm by it, to be used for preservation.
It is a kind of by using the method for the freezing seminal fluid of collecting from a male dogs of described compositions that second purpose of the present invention is to provide, can store a very long time and can being carried easily by this seminal fluid of described method, can be thawed at reasonable time by this frozen dog sperm of described method, to be used for artificial insemination.
The 3rd purpose of the present invention is to provide a kind of application to carry out test-tube method according to the refrigerated seminal fluid of freezing method, can breed excellent animal by described method, and this method can be promoted extensively.
For finishing first purpose, the invention provides a kind of compositions that is used for frozen dog sperm, it comprises 0.25-4.5g sodium citrate, 0.45-4.0g dextrose, 0.01-0.3g penicillin, 50 based on the 100ml distilled water, 000-500,000IU streptomycin, 0.25-0.45g catalase, 0.045-0.25g egg yolk, 1.5-15ml glycerol and 0.02-0.1 μ g dog serum.
For finishing second purpose, the invention provides the method for using the said composition frozen dog sperm, it may further comprise the steps: (a) collect the seminal fluid of male dogs by massage method or electric shock (electrical shock), (b) the centrifugal seminal fluid of collecting, separation is also removed supernatant, only stay the seminal fluid of lower floor, (c) seminal fluid with lower floor mixes with the compositions equal-volume, pre-cold mixt, (d) further said composition is joined in the mixture that step (c) obtains, the volume ratio of seminal fluid and said composition was reduced to 1: 2, once more the mixture that obtains of cooling, (e) liquid nitrogen is joined in the mixture that step (d) obtains freezing this mixture.
For finishing the 3rd purpose, the invention provides and be used for the test-tube method of dog, comprise the following steps: (a) detect female dog accurately the onset of ovulation to improve reproduction rate, (b) the refrigerated seminal fluid that will prepare for the method for finishing second purpose of the present invention joins in the mixture of 0.3-1.5wt% sodium chloride solution and dog serum, be heated to 30-40 ℃, it is thawed, (c) mobility of sperm in the seminal fluid that thaws by measurement microscope, (d) seminal fluid that will thaw is injected into the vagina of female dog, massages the clitoris of female dog.
The present inventor has carried out extensive studies; method that is used for frozen dog sperm and the compositions of using it have been developed; found by dog serum is combined with the compositions that is similar to the conventional diluent that is used for cattle or pig, can the stored frozen dog sperm, finished the present invention thus.
Below the present invention will be described in further detail.
The compositions that is used for frozen dog sperm is described earlier.
According to the preparation of compositions that is used for frozen dog sperm of the present invention be by will being used to regulate pooling feature, acidity (pH) and sperm osmotic pressure the 0.25-4.5g sodium citrate, as the 0.45-4.0g dextrose of metabolism and energy source, as the antibiotic 0.01-0.3g penicillin, 50 that is used to protect from infection, 000-500,000IU streptomycin, 0.25-0.45g catalase, 0.045-0.25g egg yolk, 1.5-15ml glycerol and 0.02-0.1 μ g dog serum uniform mixing carry out in the 100ml distilled water.Zhi Bei compositions is that unit is freezing with about 2ml thus, when using it is thawed.
Can store the seminal fluid a very long time according to compositions of the present invention, and can keep the survival ability and the mobility of the sperm that contains in the frozen semen simultaneously, it contains the dog serum that shows normal fertility and transfer rate after thawing.
Below, will the method that is used for frozen dog sperm that use said composition be described.
The frozen dog sperm first step is to collect dog sperm earlier.As the seminal fluid collecting method, can use method well known in the art, and without any restriction, preferred massage method or electric shock (electricalshock).Below concise and to the point most widely used massage method described.
Because sperm mixes the back with urine rapidly dead, preferred male dog elder generation before collecting seminal fluid urinates.Then the genitals of male dog is inserted in the testing tube with artificial vagina and massage, the seminal fluid of male dog is injected in the testing tube.More effective in order to make by this method collection seminal fluid, allow the female dog that oestruses around male dog, pace up and down.Perhaps, the freezing Cotton Gossypii or the sponge that will contain the vaginal secretions of heat dog thaw, and allow female dog smell its abnormal smells from the patient when collecting seminal fluid.
When using massage method collection seminal fluid, seminal fluid is divided into three duplicate samples, in different test tubes, collect.Wherein, the seminal fluid sample that contains sperm is the sample of second part of collection.The interval that every duplicate samples is collected approximately is 10-20 second.Yet, can't distinguish to such an extent as to the interval between first and second duplicate samples is too short in some cases.Preferably use second duplicate samples in the present invention.Yet, when can't all collecting in the testing tube three duplicate samples standby when second duplicate samples is distinguished first duplicate samples.
The seminal fluid that arrives by the centrifugal method separated and collected then, abandoning supernatant only stays the sedimentary seminal fluid of lower floor, and carried out 6 minutes centrifugal the branch at about 500-1100RPM/.
The sedimentary seminal fluid of lower floor is mixed at the room temperature equal-volume with the compositions of above-mentioned preparation, and pre-then cold mixt is to form the mixture of pre-cooling.Wherein be about 10-20 minute cool time, and chilling temperature is about 5 ℃.At room temperature the seminal fluid of this pre-cooling and the mixture of compositions are diluted to 1: 2 volume ratio with more compositions, and then under identical temperature and time, cool off to form cold again mixture.Through the cold again cooling step of pre-cold-peace, the complete uniform mixing of seminal fluid and mixture.
Draw the cooling mixture once more of scheduled volume then by dropper, 2-3 is dripped to the surface of dry ice.Because the surface temperature of dry ice is approximately-79 ℃, this mixture is frozen to this temperature.Should separate with dry ice by refrigerated mixture, place the container of-196 ℃ of liquid nitrogen approximately is housed, seal fully then, finished the process of frozen dog sperm thus.Because this refrigerated mixture is coccoid in liquid nitrogen, this method is called as " bead (pellet) method ".
Except above-mentioned bead method, dog sperm can be undertaken freezing by the method for using suction pipe, below concise and to the point this method of describing.
Refrigerative mixture once more is subdivided into the sample of about 0.5ml, puts into suction pipe.Every 20-30 suction pipe is divided into one group, is immersed in the container that contains liquid nitrogen.When being immersed in, suction pipe carries out in the container being that they are immersed gradually, rather than being immersed in the bottom of container immediately when freezing.The beginning, with they be immersed in container top, keep one section preset time, then they are immersed in the middle part of container, keep one section preset time, be immersed in the bottom of container then.
Refrigerated thus seminal fluid can transport and store a very long time, can be used as artificial insemination if desired after thawing.
The artificial insemination method of using frozen semen is then described below.
At first, in order to carry out artificial insemination, test to determine whether female dog is in the onset of ovulation.The test onset of ovulation adopts the technical field of the invention known method to carry out, as vaginal cell test or hormone test.
Then, by the following method with refrigerated semen thawing: refrigerated seminal fluid is joined in the mixture of a small amount of dog serum of 0.3-1.5wt% sodium chloride solution and heating, to 30-40 ℃, it helps keeping the survival ability and the shape of sperm with mixture heated.And although this mixture can direct heating, owing to be difficult to the control temperature, the sealed container that preferably will contain sodium chloride solution and dog serum is immersed in about 50 ℃ of hot water and heats.
Before using the seminal fluid thaw thus and carrying out artificial insemination, quantity, shape and the mobility of sperm in the seminal fluid that thaws by measurement microscope.The quantity of the sperm that moves forward is depended in successful artificial insemination.Mobility according to them is divided into four classes with sperm.
0 class represents that sperm is that afterbody moves, and the situation that can not move forward, 1 class represents to be less than the situation that 20% sperm moves forward, 2 classes are represented the situation that the sperm of 20-50% moves forward, 3 classes are represented the situation that the sperm of 50-80% moves forward, and 4 classes represent to surpass the situation that 80% sperm moves forward.When the seminal fluid that will thaw places on the slide and during by microscopic examination, has only the seminal fluid of 2 classes or higher class mobility can be used for artificial insemination.
Then, the seminal fluid that will thaw is injected into the vagina of the female dog that is in the egg-laying season, massages the clitoris of female dog, finishes artificial insemination.
Except above-mentioned artificial insemination method, the seminal fluid that thaws can be injected into the uterus of female dog by operating mode.In other words, an about 2cm osculum is cut in the umbilicus bottom, and the pull-up uterus directly is injected into the seminal fluid of male dog in the uterus of female dog by syringe.Then, peritoneal suture, abdominal muscle and skin successively.
The preferred forms of inventing
Further specify the present invention by following embodiment, they never are in order to limit the scope of the invention.
Embodiment 1
The preparation of compositions of frozen dog sperm
The application mix device will be based on the mixture uniform mixing of the following component of 100ml distilled water, with preparation frozen dog sperm compositions.
Sodium citrate 0.35g
Dextrose 2.5g
Penicillin 0.02g
Streptomycin 100,000IU
Catalase 0.35g
Egg yolk 0.05g
Glycerol 2ml
Dog serum 0.05 μ g
The freezing method of dog sperm
Collect the seminal fluid of 6 years old shepherd dog of body weight 43kg by electric shock (electrical shock).The semen volume of collecting is 6.2ml.The seminal fluid that arrives by the centrifugal method separated and collected then, abandoning supernatant only stays the sedimentary seminal fluid of lower floor, under 500-1100RPM/ divides centrifugal 6 minutes.
Isolating lower floor seminal fluid is thus mixed pre-then cold mixt at the room temperature equal-volume with the compositions of above-mentioned preparation.Wherein be 10-20 minute cool time, and chilling temperature is about 5 ℃.Then compositions is further joined in the mixture of pre-cooling, seminal fluid is mixed with volume ratio with compositions at 1: 2.Then the mixture that obtains is cooled off under identical temperature and time once more.
Draw the refrigerative once more mixture of scheduled volume then by dropper, 2-3 is dripped to the surface of dry ice.Because the surface temperature of dry ice is-79 ℃ approximately, this mixture is frozen to this temperature.Should separate with dry ice by refrigerated mixture, place the container of-196 ℃ of liquid nitrogen approximately is housed, seal fully then, finished the process of frozen dog sperm thus.
Use the artificial insemination of frozen semen
3 years old female shepherd dog of three individual weight 30kg is as artificial insemination.Whether be in the onset of ovulation by the vaginal cell test with definite female dog.
Then, need be with the refrigerated semen thawing of said method.Thaw and be to be undertaken: refrigerated seminal fluid is joined in the mixture of 0.3-1.5wt% sodium chloride solution and a small amount of dog serum,, kept 5 hours mixture heated to 38 ℃ by following method.
Then, before using this seminal fluid that thaws and carrying out artificial insemination, place by the seminal fluid that will thaw and use microscopic examination on the slide, to detect the mobility of the sperm of surviving.The quantity of the sperm that moves forward is depended in successful artificial insemination.The percentage ratio of the sperm that moves forward is approximately 85%, and it is suitable for artificial insemination.
Next step, the seminal fluid that will thaw is divided into three parts, is injected into the vagina of the female dog that is in the egg-laying season then.Massage the clitoris 5 minutes of every female dog then, finish artificial insemination.
Carried out the pregnancy detection in 1 month after artificial insemination is finished, all three female dogs are all conceived as a result.Three female dogs bore 4,6 and 9 healthy doggies respectively afterwards.
Embodiment 2
In the present embodiment, prepare the compositions of frozen dog sperm according to the method identical with embodiment 1, what only be to use is 0.02 μ g dog serum.
By the method identical, use said composition and carry out the freezing and artificial insemination of dog sperm then with embodiment 1.
In this embodiment, use 5 years old male shepherd dog of a body weight 40kg and three body weight of 3 years old female shepherd dog as 25kg, 30kg and 28kg.
The mobility test result of seminal fluid of thawing shows that the percentage ratio that sperm moves forward is 60%, and it is suitable for artificial insemination.
Carried out the pregnancy detection in 1 month after artificial insemination is finished, two female dogs of result are all conceived.Two female dogs bore 5 and 6 healthy doggies respectively afterwards.
Comparative examples 1
In this comparative examples, be used for the same combination of frozen dog sperm according to embodiment 1 preparation, and according to the method frozen dog sperm identical, except the volume ratio of compositions and dog sperm is 1: 1 with embodiment.And collect the dog sperm of same male dog according to the method identical with embodiment 1.
According to the method identical refrigerated seminal fluid that thaws, the mobility of sperm in the seminal fluid that test is thawed with embodiment 1.Test result shows percentage ratio that sperm moves forward less than 20%, and it is not suitable for artificial insemination.
Comparative examples 2
In this comparative examples, prepare the same combination that is used for frozen dog sperm according to the mode identical with embodiment 1, in compositions, do not contain the dog serum.Identical among freezing method and defreezing method and the embodiment 1, and collect the dog sperm of same male dog according to the method identical with embodiment 1.
The mobility test shows of and the sperm that thaw freezing according to this comparative examples, sperm is the afterbody activity, and can not move forward, and it is not suitable for artificial insemination.
As mentioned above, the invention provides the method that is used for frozen dog sperm, wherein dog sperm is because the characteristic of its sperm is difficult to stored frozen, and the present invention also provides the compositions of using in this method.Therefore, dog sperm can be preserved under freezing state, can keep the improved seeds of dog thus.And refrigerated male dogs seminal fluid can be used for artificial insemination at reasonable time and place, therefore can increase the natality of dog.
Claims (2)
1. compositions that is used for frozen dog sperm, it comprises 0.25-4.5g sodium citrate, 0.45-4.0g dextrose, 0.01-0.3g penicillin, 50 based on the 100ml distilled water, 000-500,000IU streptomycin, 0.25-0.45g catalase, 0.045-0.25g egg yolk, 1.5-15ml glycerol and 0.02-0.1 μ g dog serum.
2. the method for application such as the desired compositions frozen dog sperm of claim 1, it may further comprise the steps:
(a) collect the seminal fluid of male dogs by massage method or electric shock;
(b) the centrifugal seminal fluid of collecting separates and removes supernatant, only stays the seminal fluid of lower floor;
(c) seminal fluid with lower floor mixes with the said composition equal-volume, pre-cold mixt;
(d) further said composition is joined in the mixture that step (c) obtains, the volume ratio of seminal fluid and said composition was reduced to 1: 2, once more the mixture that obtains of cooling; With
(e) liquid nitrogen is joined in the mixture that step (d) obtains freezing this mixture.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1019990015147A KR19990078560A (en) | 1999-04-28 | 1999-04-28 | freezing method of dog sperm and artificial insemination method utilizing the sperm and compound for freezing the sperm thereof |
| KR1999/15147 | 1999-04-28 | ||
| KR2000/21012 | 2000-04-20 | ||
| KR1020000021012A KR100320588B1 (en) | 1999-04-28 | 2000-04-20 | Compositions for freezing dog sperm, freezing methods of the dog sperm utilizing the compositions and artificial insemination method empolying the freezed dog sperm |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1351484A CN1351484A (en) | 2002-05-29 |
| CN1170511C true CN1170511C (en) | 2004-10-13 |
Family
ID=26635013
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB008080003A Expired - Fee Related CN1170511C (en) | 1999-04-28 | 2000-04-26 | Compositions for freezing dog sperm, method of freezing the dog sperm utilizing the compositions and artificial insemination method employing the frozen dog sperm |
Country Status (7)
| Country | Link |
|---|---|
| JP (1) | JP2002543101A (en) |
| KR (1) | KR100320588B1 (en) |
| CN (1) | CN1170511C (en) |
| AU (1) | AU773578B2 (en) |
| DE (1) | DE10084534B4 (en) |
| GB (1) | GB2364233B (en) |
| WO (1) | WO2000066028A1 (en) |
Families Citing this family (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100433280B1 (en) * | 2000-09-22 | 2004-05-31 | 대한민국 | Diluent for Boar Sperm Freezing |
| KR20030014891A (en) * | 2001-08-13 | 2003-02-20 | (주)베티즌 | Compositions for cooling and cryopreservation of canine sperm, Methods for cooling and cryopreservation using the compositions, and Canine Articificial Insemination |
| CN1331390C (en) * | 2003-12-24 | 2007-08-15 | 甘孜州家畜改良站 | Method for making frozen spermatozoon of yak |
| CN100405902C (en) * | 2004-02-20 | 2008-07-30 | 辽宁省辽宁绒山羊原种场 | Frozen semen of animal and its making method |
| CN100563665C (en) * | 2006-09-11 | 2009-12-02 | 华中农业大学 | A kind of cow sex-control sperm freezing preservation seminal fluid dilution |
| KR101064415B1 (en) * | 2010-04-30 | 2011-09-14 | (주)미래생명공학연구소 | Method for treatment of spermatozoa in external fertilization of domestic animal |
| CN102379279A (en) * | 2011-11-17 | 2012-03-21 | 张大宇 | Refrigerating fluid for storing Tibetan spaniel sperm |
| CN103141471B (en) * | 2013-02-01 | 2015-09-30 | 四川农业大学 | A kind of Kunming hangquan semen freezing formula of liquid and compound method |
| CN103900884B (en) * | 2014-04-11 | 2016-09-14 | 深圳市博锐德生物科技有限公司 | Refining is combined quality-control product and preparation method thereof, test kit |
| CN104758084A (en) * | 2015-04-20 | 2015-07-08 | 陈绪凯 | Method for artificial insemination for cow through sex control and semen freezing |
| JP7033604B2 (en) * | 2016-10-18 | 2022-03-10 | スマートボウ・ゲーエムベーハー | Equipment for artificial insemination of mammals |
| CN107251892A (en) * | 2017-05-25 | 2017-10-17 | 中国热带农业科学院热带作物品种资源研究所 | It is a kind of suitable for the solution for dilution of cock semen of torrid areas, preparation and its application process |
| ES2695248A1 (en) * | 2017-06-26 | 2019-01-02 | Spermbiobank Slu | METHOD AND KIT FOR FREEZING AND TRANSPORTATION OF SEMEN (Machine-translation by Google Translate, not legally binding) |
| CN107494520B (en) * | 2017-09-25 | 2020-07-28 | 江苏农牧科技职业学院 | Dog semen freezing diluent and freezing method |
| KR102536863B1 (en) | 2019-11-25 | 2023-05-30 | 서울대학교산학협력단 | A Composition for semen cryoprotectant of animal including amniotic membrane derived stem cell-conditioned media and method of making thereof |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SU587934A1 (en) * | 1973-07-19 | 1978-01-15 | Украинский Научно-Исследовательский Институт Разведения И Искусственного Осеменения Крупного Рогатого Скота | Animal sperm dilutant |
| US4007087A (en) * | 1975-10-17 | 1977-02-08 | Gametrics Limited | Sperm fractionation and storage |
| SE7800809L (en) * | 1977-01-26 | 1978-07-27 | Fisons Ltd | PROCEDURE FOR THE PREPARATION OF NEW SALTS OF BENZOPYRANE DERIVATIVES |
| JP3586797B2 (en) * | 1994-07-11 | 2004-11-10 | 独立行政法人理化学研究所 | Sperm cryoprotectant |
| KR0160090B1 (en) * | 1995-05-01 | 1999-02-18 | 김광희 | Artificial insemination diluent for liquid sperm of pig |
-
2000
- 2000-04-20 KR KR1020000021012A patent/KR100320588B1/en not_active IP Right Cessation
- 2000-04-26 WO PCT/KR2000/000396 patent/WO2000066028A1/en active Application Filing
- 2000-04-26 GB GB0126189A patent/GB2364233B/en not_active Expired - Fee Related
- 2000-04-26 JP JP2000614915A patent/JP2002543101A/en active Pending
- 2000-04-26 CN CNB008080003A patent/CN1170511C/en not_active Expired - Fee Related
- 2000-04-26 AU AU44351/00A patent/AU773578B2/en not_active Ceased
- 2000-04-26 DE DE10084534T patent/DE10084534B4/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| AU773578B2 (en) | 2004-05-27 |
| AU4435100A (en) | 2000-11-17 |
| CN1351484A (en) | 2002-05-29 |
| DE10084534T1 (en) | 2002-05-02 |
| KR100320588B1 (en) | 2002-01-12 |
| JP2002543101A (en) | 2002-12-17 |
| WO2000066028A1 (en) | 2000-11-09 |
| KR20000049647A (en) | 2000-08-05 |
| GB0126189D0 (en) | 2002-01-02 |
| GB2364233B (en) | 2003-06-25 |
| DE10084534B4 (en) | 2007-09-13 |
| GB2364233A (en) | 2002-01-23 |
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