JP5623895B2 - Egg collection and embryo transfer equipment - Google Patents

Description

  The present invention relates to an egg collection and embryo transfer device for animals.

  For genetic modification, production of chimeric animals, artificial growth of animals, etc., eggs and embryos are collected from the uterus of the animal, processed as necessary, and then transferred to the uterus of another individual. There is a need.

  In order to collect fertilized eggs and embryo transfer, laparotomy has been required so far, and the method using laparotomy has also invaded animals, and adhesion of the surgical site has also been a problem.

  A non-surgical embryo transfer method has also been reported instead of a transfer method by laparotomy (see Non-Patent Documents 1 and 2). In this method, a stainless steel transplant catheter is used, and there is a problem that the catheter is thick and difficult to enter the uterine ostium. there were. Furthermore, in this method, an otoscope (ear endoscope) was used to find the uterine ostium. However, in the method using the otoscope, it is difficult to find the uterine ostium, and the method has not been widespread.

Thomson JA., Et al., J Med Primatol 1994: 23: 333-336 Marshall VS., et al., J Med Primatol 1997: 26: 241-247

  The present invention can collect a fertilized egg safely and easily from a small animal such as a small monkey, and can further safely and easily and efficiently transfer an embryo. The purpose is to provide equipment.

  In 2008, an ultra-fine endoscope for animals (TESARA, AVS Co., Ltd.) was developed, which enabled operation while looking at the uterine ostium. Therefore, in order to enable non-surgical transplantation with almost no invasion to animals, the present inventor has developed an egg collection and embryo transfer device that easily enters the uterine ostium and does not damage the uterine ostium. As a result, the use of a double-structured catheter consisting of a thin 25-gauge stainless steel needle to find the uterine ostium and a 18-gauge catheter made of a soft material such as tapered Teflon is impossible. The present inventors have found that the egg collection and embryo transfer device can be easily inserted into the uterine ostium without exerting excessive force. By using the egg collection and embryo transfer device, it was possible to collect eggs from the uterus and transplant the embryo into the uterus without damaging the uterine ostium of the animal. In addition, it is difficult to insert an 18-gauge catheter directly into the uterine ostium, but by using the above-described egg collection and embryo transfer device having a double structure, an 18-gauge catheter can be easily inserted into the uterine ostium. Could be inserted into. Further, by producing the above 18 gauge catheter with a transparent material, the amount of embryos and medium to be transplanted could be visually observed, and the transplanting operation could be facilitated.

That is, the present invention is as follows.
[1] An inner core made of a metal needle; a catheter made of a meridian tube having a distal end and a proximal end made of a flexible material; a flexible material into which the inner core or the catheter can be slidably inserted. A guide catheter having a distal end and a proximal end and having a tapered tip; and the guide catheter or a combination of the guide catheter and the inner core or the catheter can be inserted. Including a guide tube made of a hard material as a component, inserted into the uterine ostium with the inner core inserted into the guide catheter, then removed and used to insert the catheter. A small monkey egg collection and embryo transfer device that is longer than the guide catheter and longer than the guide tube.
[2] Inner core needle size is 24-26 gauge, length is 100-200mm, catheter size is 22-24 gauge, tube length is 100-200mm, guide catheter size is 18 The instrument for egg transplantation and embryo transfer according to [1], which is ˜20 gauge, the length of the tube portion is 70 to 130 mm, the guide tube size is 3 to 10 mm in outer diameter, and the length is 50 to 100 mm.
[3] The egg collection and embryo transfer device according to [1] or [2], further including a syringe.
[4] A method for collecting a fertilized egg from the uterus of a small monkey using the egg collection and embryo transfer device according to any one of [1] to [3],
Inserting a guide tube into the vagina from the vaginal opening of a small monkey;
An inner core is inserted into the guide catheter to form a double-structure catheter, and the double-structure catheter is inserted through the guide tube until the inner core and the distal end of the guide catheter reach the uterus;
After insertion, removing the inner core from the guide catheter;
Inserting a catheter into the guide catheter;
Flowing perfusate into the uterus through an opening at the proximal end of the catheter;
The perfusate that enters the guide catheter from the opening at the distal end of the guide catheter, passes through the guide catheter, and flows out of the body from the opening at the proximal end of the guide catheter is collected, and fertilization contained in the perfusate A method of collecting a fertilized egg from the uterus of a small monkey, including collecting an egg.
[5] A method for transplanting an embryo into the uterus of a small monkey using the egg collection and embryo transfer device according to any one of [1] to [3],
Inserting a guide tube into the vagina from the vaginal opening of a small monkey;
An inner core is inserted into the guide catheter to form a double-structure catheter, and the double-structure catheter is inserted through the guide tube until the inner core and the distal end of the guide catheter reach the uterus;
After insertion, removing the inner core from the guide catheter;
Inserting a catheter into the guide catheter;
A method for implanting an embryo into a small monkey uterus, comprising injecting a liquid containing the embryo into the uterus through the catheter.

  The egg collecting / embryo transfer device of the present invention is inserted into the uterine ostium as a catheter having a double structure in which a rigid metal inner core and a guide catheter having flexibility and taper are combined. By using a rigid inner core and a flexible and tapered guide catheter in combination, the device of the present invention can be easily inserted into the uterine ostium without damaging the uterine ostium. By replacing the inner core with a catheter after insertion into the uterine ostium, fertilized eggs can be collected from the uterus by perfusing perfusate into the uterus through the catheter, and the embryo is transplanted into the uterus through the catheter Can do. By using the egg collection and embryo transfer device of the present invention, the egg collection and embryo transfer of animals such as small monkeys can be performed non-invasively, non-invasively and efficiently to animals. Egg production and embryo transfer for artificial growth of animals and embryo transfer can be repeated safely, easily and efficiently in the same individual, so that an economic effect can be obtained and the welfare of animals can be improved.

It is a figure of the inner core which comprises the instrument for egg collection and embryo transfer of this invention. It is a figure of the catheter which comprises the instrument for egg collection and embryo transfer of this invention. It is a figure of the guide catheter which comprises the instrument for egg collection and embryo transfer of this invention. It is a figure of the guide tube which comprises the instrument for egg collection and embryo transfer of this invention. It is a figure which shows the state which combined the inner core, the guide catheter, and guide tube which comprise the instrument for egg collection and embryo transfer of this invention. It is a figure which shows the state which combined the guide catheter and guide tube which comprise the instrument for egg collection and embryo transfer of this invention. It is a figure which shows the state which combined the catheter, guide catheter, and guide tube which comprise the instrument for egg collection and embryo transfer of this invention. It is a figure which shows the outline | summary of the method of extract | collecting an egg using the instrument for egg collection and embryo transfer of this invention. It is a figure which shows the outline | summary of the method of transplanting an embryo using the instrument for egg collection and embryo transfer of this invention. Image observed by ultrasound when performing transvaginal embryo transfer using the egg collection and embryo transfer device of the present invention (FIG. 10A), ultrasound observation image of uterus 4 weeks after embryo transfer (FIG. 10B left) ) And an ultrasonic observation image (right of FIG. 10B) of the uterus not transplanted with an embryo. It is a figure which shows the implantation rate at the time of transplanting by the transvaginal embryo transfer method and the laparotomy embryo transfer method. It is a figure which shows the comparison of a laparotomy embryo transfer method and a transvaginal embryo transfer method. It is a figure which shows the egg collection efficiency of a laparotomy egg collection method, a percutaneous egg collection method, and the transvaginal egg collection method of this invention. It is a figure which shows the comparison of the laparotomy egg collection method, the percutaneous egg collection method, and the transvaginal egg collection method of this invention.

  Hereinafter, the present invention will be described in detail.

  The present invention relates to an egg collection and embryo transfer device for collecting a fertilized egg from an animal uterus non-invasively, transvaginally, or transplanting an embryo into the uterus without surgical operation such as laparotomy. In this method, an egg is collected using the instrument and an embryo is transplanted. Here, the embryo to be transplanted refers to an early embryo in animal ontogeny, and includes embryos in the blastocyst stage before fertilization to implantation.

  The animal which is the subject of the present invention is a relatively small mammal, preferably a small primate, for example, an average female body weight of 250 to 5000 g or less, or an adult female body length of 18 to 100 cm or less. It is a primate. Specifically, common marmoset (Callithrix jacchus), pygmy marmoset (Cebuella pygmaea), cottonbosch panchee (Satuinus oedipus), acatetamarin (Sauinaus midas), small lion monkey belonging to the marmoset family such as Golden Lion Tamarin (Leontopithecus rosalia); common Small monkeys belonging to the squirrel monkey subfamily such as squirrel monkey (Saimiri sciureus); small monkeys belonging to the galagoid family such as Galago senagalensis; primates belonging to the Loris family such as Slow loris (Nycticebus coucang) Primates belonging to the family Aiaidae; rhesus monkey (Macaca mulatta); cynomolgus monkey (Macaca fascicularis).

  In the method of the present invention, an embryo transfer device is used. These instruments can be easily inserted into the uterine ostium without damaging the uterine ostium of an animal, and a fertilized egg can be easily collected from the uterus or an embryo can be transplanted into the uterus. Since the embryo transfer device can also be used for collecting eggs, the device of the present invention is called an egg collection and embryo transfer device.

  The egg collection and embryo transfer instrument of the present invention has good operability and does not damage the vagina or cervix when inserted through the vagina or uterine ostium.

  Hereinafter, the egg collection and embryo transfer device of the present invention will be described with reference to the drawings.

  The egg collecting and embryo transfer device of the present invention includes an inner core 1 made of a metal needle (FIG. 1); a catheter 4 made of a meridian tube having a distal end and a proximal end made of a flexible material (FIG. 2); A guide catheter 7 (FIG. 3) having a distal end and a proximal end, which is made of a flexible material and has elasticity, into which the inner core or the catheter can be slidably inserted and removably inserted; and a guide A guide tube 10 (FIG. 4) made of a hard material into which a catheter or a combination of the catheter and the inner core 1 or the catheter 4 can be inserted is included as a component.

  The inner core 1 shown in FIG. 1 is inserted into a guide catheter and used as a catheter having a double structure. Thus, when the inner core 1 is inserted into the guide catheter and used as a double-structure catheter, the inner core 1 imparts rigidity to the guide catheter so that it is difficult to bend when the guide catheter is inserted into the uterine ostium. Moreover, the inner core 1 has rigidity, and can give rigidity to the whole double structure catheter, and can be easily inserted into the uterine ostium. On the other hand, although the inner core 1 is thin and difficult to insert into the uterine ostium, it is inserted into the uterine ostium by combining it with a guide catheter 7 having a certain thickness and flexibility to form a double structure catheter. Becomes easier. As a result, the guide catheter 7 with the inner core 1 inserted can be inserted into the uterine ostium. That is, the inner core 1 functions as a guide wire for inserting the guide catheter 7 into the uterine ostium. The inner core 1 is preferably made of a metal such as stainless steel, a Co—Cr alloy, or a titanium alloy that has a small biological reaction and is biocompatible. The inner core 1 may or may not be hollow. The shape of the tip is not limited, but it is desirable that the tip is rounded so as not to damage the vagina or the uterus when inserted into the uterus through the vagina. The inner core 1 has a length such that the distal end protrudes from the distal end of the guide catheter 7 when inserted into the guide catheter 7. The inner core 1 has a grip portion 3 for gripping with a finger on the base. In the present invention, the distal end portion refers to the side inserted into the uterine ostium, and the base portion refers to the end portion that is outside the body during egg collection or embryo transfer operation, and is the side held by the fingers. The grip portion 3 may be manufactured integrally with the metal needle portion 2, or may be manufactured separately from and combined with the needle portion 2. When manufacturing separately, the needle part 2 and the grip part 3 may be joined by bonding, welding, fitting, or the like.

  The catheter 4 shown in FIG. 2 is a tube having an opening at the distal end and the base, and can inject perfusate for collecting eggs from the base, which serves as a passage for the perfusate. Although the catheter 4 is thin and difficult to insert into the uterine ostium, it can be easily inserted into the uterine ostium by combining it with a guide catheter 7 having a certain thickness and flexibility to form a double structure catheter. Become. The catheter 4 is inserted into the uterus by removing the inner core 1 from the guide catheter 7 inserted into the uterus with the inner core 1 inserted and then inserting it into the guide catheter 7. The catheter 4 has a length such that the distal end protrudes from the distal end of the guide catheter 7 when inserted into the guide catheter 7. The catheter 4 includes a tube portion 5 and a connection portion 6 for connecting to a syringe or the like at the base portion. The connecting portion 6 also has a function as a gripping portion for gripping with fingers. The catheter 4 is made of a biocompatible material. Polyester; Polyolefin such as polyethylene, polypropylene, ethylene-propylene copolymer; Polyamide such as 6 nylon and 66 nylon; Polyester such as polyethylene terephthalate and polybutylene terephthalate; Polyacetal; Polyphenylene sulfide; Polyetheretherketone: polytetrafluoroethylene (PTFE), tetrafluoroethylene / perfluoroalkyl vinyl ether copolymer (PFA), tetrafluoroethylene / hexafluoropropylene copolymer (4.6 fluoride) (FEP), tetra Fluoroethylene / ethylene copolymer (ETFE), polyvinylidene fluoride (difluoride) (PVDF), polychlorotrifluoroethylene (trifluoride) (PCTFE), chlorotrifluoroethylene Ren copolymer (ECTFE) include fluorine resins such as, PTFE Among, PFQ, Teflon FEP, etc. (registered trademark) can be suitably used. A material obtained by coating a material other than the above fluororesin with a fluororesin can also be used.

  The guide catheter 7 shown in FIG. 3A is a tube having an opening at the tip, and is inserted into the uterine ostium with the inner core 1 inserted, and the inner core 1 is removed in the inserted state. The catheter 4 is inserted. The guide catheter 7 has a role as a guide when the catheter 4 is inserted into the uterus, and is thicker and less bent than the catheter 4. Further, the guide catheter 7 has a tapered tip (FIG. 3B) and a tapered shape so as not to damage the uterus. Furthermore, it is made of a flexible material and has elasticity. The term “flexibility” refers to a property that is flexible and does not break even when bent. The guide catheter 7 has a connection portion 9 for connecting the tube portion 8 and a syringe or the like. The connecting portion 9 also has a function as a gripping portion for gripping with fingers. The guide catheter 7 is made of a biocompatible material, polyester; polyolefin such as polyethylene, polypropylene and ethylene-propylene copolymer; polyamide such as 6 nylon and 66 nylon; polyester such as polyethylene terephthalate and polybutylene terephthalate; polyacetal; polyphenylene sulfide. Polyether ether ketone; polytetrafluoroethylene (PTFE), tetrafluoroethylene / perfluoroalkyl vinyl ether copolymer (PFA), tetrafluoroethylene / hexafluoropropylene copolymer (4.6 fluoride) (FEP), Tetrafluoroethylene / ethylene copolymer (ETFE), polyvinylidene fluoride (difluoride) (PVDF), polychlorotrifluoroethylene (trifluoride) (PCTFE), chlorotrifluoroethylene · Ethylene copolymer (ECTFE) include fluorine resins such as, PTFE Among, PFQ, Teflon FEP, etc. (registered trademark) it can be suitably used. A material obtained by coating a material other than the above fluororesin with a fluororesin can also be used. The guide catheter 7 is preferably transparent or translucent. In the case of being transparent or translucent, the amount of fertilized egg to be collected, the embryo to be transplanted and the amount of the medium can be visually observed, and the egg collecting and embryo transfer operations can be easily and reliably performed.

  The guide tube 10 made of a hard material into which the guide catheter 7 can be inserted is a tube having both ends opened. The guide tube 10 is inserted from the vagina. During the egg collection, the base opening is outside the body, and the tip opening is The guide catheter 7 and the catheter 4 are inserted into the uterus through the guide tube while being held in the vagina. The guide tube 10 preferably has a tapered shape with a tapered tip so as not to damage the vagina. Since the guide tube is inserted into the vagina, the guide tube is made of a hard material. Examples of the hard material include glass; metals such as biocompatible stainless steel, Co-Cr alloy, and titanium alloy.

  The size of the components of the egg collection and embryo transfer instrument according to the present invention depends on the size of the target animal. For example, in the case of a common marmoset with a female body length of 20 to 25 cm, it is as follows. The following egg collection and embryo transfer devices can be widely used for small monkeys of the same size as common marmoset. What is necessary is just to change a size suitably according to the magnitude | size of a monkey with respect to the other small monkey from which a magnitude | size differs.

  The size of the needle portion 2 of the inner core 1 is 24 to 26 gauge, and the length is 100 to 200 mm, preferably 125 to 175 mm, more preferably 140 to 160 mm, not including the grip portion 3.

  The size of the catheter 4 is 22 to 24 gauge, and the length is 100 to 200 mm, preferably 135 to 185 mm, more preferably 150 to 170 mm, in terms of the length of the tube portion 5 not including the connecting portion (gripping portion) 6.

  The size of the guide catheter 7 is 18 to 20 gauge, the distal end is tapered in a tapered shape, and the outer diameter of the most distal end is 0.3 to 0.4 mm smaller than the outer diameter of the base. The length is 80 to 140 mm, preferably 100 to 120 mm, as the length of the tube portion 8 not including the connecting portion (gripping portion) 9.

  The guide tube 10 has an outer diameter of 3 to 10 mm and a length of 50 to 100 mm, preferably 70 to 80 mm.

  The length of the inner core 1 and the length of the catheter 4 are longer than the length of the guide catheter 7, and when the inner core 1 or the catheter 4 is inserted into the guide catheter 7 to the base, the distal end portion 10 of the inner core 1 and the catheter 4. ˜100 mm, preferably 25 to 75 mm, protrudes from the distal end portion of the guide catheter 7. Further, the length of the guide tube 10 is shorter than the lengths of the inner core 1, the catheter 4 and the guide catheter 7, and when the inner core 1, the catheter 4 or the guide catheter 7 is inserted into the guide tube 10, the distal end portion and the base portion thereof are arranged. 10 to 50 mm, projecting from the tip and base of the guide tube, respectively.

  Further, the outer diameters of the inner core 1 and the catheter 4 are made smaller than the inner diameter of the guide catheter 7 so that the inner core 1 and the catheter 4 are slidably inserted into the guide catheter 7. In particular, when an egg is collected, an egg in the uterus is collected through the guide catheter 7 with the catheter 4 inserted into the guide catheter 7, so that the egg passes when the catheter 4 is inserted into the guide catheter 7. There needs to be space available.

  A fertilized egg is collected from the uterus using the egg collection and embryo transfer device of the present invention as follows.

1 Insert the guide tube 10 into the vagina from the vaginal opening 12 of the subject animal. At this time, the base portion of the guide tube protrudes outside the body, and the distal end portion is located in the vagina.

2 Insert the inner core 1 into the guide catheter 7 to obtain a double-structure catheter. At this time, the distal end portion of the inner core 1 protrudes from the distal end portion of the guide catheter 7.

3. Insert the guide catheter 7 with the inner core 1 inserted through the guide tube 10 until the inner core 1 and the distal end of the guide catheter 7 reach the uterus. At this time, an endoscope may be inserted into the guide rod 10, the uterine ostium may be found from the endoscopic image, and the inner core 1 and the guide catheter 7 may be inserted into the uterine ostium. Since the inner core 1 is made of metal and has rigidity, the guide catheter 7, which is a thin flexible hollow tube, can be inserted into the uterus using the rigidity. The state of the guide tube 10, the guide catheter 7, and the inner core 1 at this time is shown in FIG.

4 Pull out the inner core 1 from the guide catheter 7. At this time, the distal end of the guide catheter 7 is inserted into the uterus. The state of the guide tube 10 and the guide catheter 7 at this time is shown in FIG.

5 Insert the catheter 4 into the guide catheter 7. The catheter 4 is inserted into the uterus using the guide catheter 7 as a guide, and is held in a state where the distal end portion is located in the uterus and the proximal end portion is located outside the body. At this time, in the uterus, the catheter 4 is inserted into the guide catheter 7 and the distal end portion of the catheter 4 protrudes from the distal end portion of the guide catheter 7. Further, even outside the body, the proximal end portion of the catheter 4 protrudes from the proximal end portion of the guide catheter 7. The state at this time is shown in FIG.

6 Flow the perfusate into the uterus from the opening at the proximal end of the catheter 4. At this time, a syringe filled with perfusate may be connected to a connection portion provided at the proximal end of the catheter 4, and the perfusate in the syringe may be poured into the uterus, or exists at the proximal end of the catheter 4. An appropriate tube may be connected to the connection portion, and the perfusate may be poured into the uterus through the tube.

7 The perfusate flows into the uterus from the distal end of the catheter, and the perfusate that has flowed out perfuses the uterus. The perfusate enters the guide catheter 7 through the opening of the distal end of the guide catheter 7 and passes through the guide catheter 7. Then, it flows out of the body from the opening at the proximal end of the guide catheter 7. At this time, since the egg existing in the uterus comes out of the body through the guide catheter 7 together with the perfusate, the egg may be collected and collected.

  FIG. 8 shows an outline of the egg collecting method. In FIG. 8, the arrow indicates the flow of the perfusate. As shown in FIG. 8, a guide catheter 7 having a catheter 4 inserted therein is inserted into a guide tube 10 inserted into the vagina 14 from the vaginal opening 12, and the distal end portions of the guide catheter 7 and the catheter 4 are in the uterine cavity 13. Located in. The perfusate is injected from the base opening of the catheter 4, and the perfusate flows into the uterine cavity 13 from the distal end opening of the catheter 4. The perfusate guides the fertilized egg 15 in the uterus into the guide catheter 7 and then the fertilized egg 15. Can pass through the guide catheter 7 and be collected from the base opening of the guide catheter 7.

  The egg collection and embryo transfer device of the present invention is used as follows to transfer an embryo into the uterus.

1 Insert the guide tube 10 into the vagina from the vaginal opening 12 of the subject animal. At this time, the base portion of the guide tube protrudes outside the body, and the distal end portion is located in the vagina.

2 Insert the inner core 1 into the guide catheter 7. At this time, the distal end portion of the inner core 1 protrudes from the distal end portion of the guide catheter 7.

3. Insert the guide catheter 7 with the inner core 1 inserted through the guide tube 10 until the inner core 1 and the distal end of the guide catheter 7 reach the uterus. Since the inner core 1 is made of metal and has rigidity, the guide catheter 7, which is a thin flexible hollow tube, can be inserted into the uterus using the rigidity. The state of the guide tube 10, the guide catheter 7, and the inner core 1 at this time is shown in FIG.

4 Pull out the inner core 1 from the guide catheter 7. At this time, the distal end of the guide catheter 7 is inserted into the uterus. The state of the guide tube 10 and the guide catheter 7 at this time is shown in FIG.

5 Insert the catheter 4 into the guide catheter 7. The catheter 4 is inserted into the uterus using the guide catheter 7 as a guide, and is held in a state where the distal end portion is located in the uterus and the proximal end portion is located outside the body. At this time, in the uterus, the catheter 4 is inserted into the guide catheter 7 and the distal end portion of the catheter 4 protrudes from the distal end portion of the guide catheter 7. Further, even outside the body, the proximal end portion of the catheter 4 protrudes from the proximal end portion of the guide catheter 7. The state at this time is shown in FIG.

6 Connect the syringe containing the liquid containing the embryo to the connection part of the catheter 4. The embryo in the syringe is transplanted into the uterus through the catheter 4.

  FIG. 9 shows an outline of the embryo transfer method. As shown in FIG. 9, a guide catheter 7 in which a catheter 4 is inserted is inserted into a guide tube 10 inserted into the vagina 14 from the vaginal opening 12, and the distal ends of the guide catheter 7 and the catheter 4 are in the uterine cavity 13. Located in. The liquid containing the embryo is injected from the base opening of the catheter 4, and the embryo is transplanted into the uterine cavity 13.

  Egg harvesting and embryo transfer equipment should be sterilized before use. Sterilization can be performed by a known sterilization method such as a high-pressure steam sterilization method, an ethylene oxide gas (EOG) sterilization method, or a radiation sterilization method.

  The present invention will be specifically described by the following examples, but the present invention is not limited to these examples.

Example 1 Embryo Transfer by Non-invasive Embryo Transfer Method Using the Egg Collection and Embryo Transfer Device of the Present Invention The embryo used was a fertilized egg collected using the egg collection device of the present invention. In addition, as a borrowed marmoset, an individual who was 2 years or older and within 10 days from the start of the luteal phase was used.
The above embryos were transplanted into a borrowed marmoset.

  Transplantation is performed by non-invasively transvaginally using the egg collection and embryo transfer device of the present invention (transvaginal embryo transfer method), and a laparoscopic embryo transfer that requires surgical laparotomy as a control Done by law.

  FIG. 10A shows an image observed by ultrasound when performing transvaginal embryo transfer using the egg collection and embryo transfer device of the present invention. FIG. 10 shows a state where the catheter guide 7 and the catheter 4 are located in the uterus. FIG. 10B shows an ultrasonic diagnostic image in the uterus 4 weeks after embryo transfer, and FIG. 10B right shows an ultrasonic diagnostic image of the uterus that was not transplanted. In the left of FIG. 10B, the open space of the uterine cavity is observed.

  FIG. 11 shows the implantation rate when transplantation is performed by the transvaginal embryo transfer method and the open embryo transfer method using the egg collection and embryo transfer device of the present invention. Non-invasive transvaginal embryo transfer resulted in an implantation rate equal to or better than open embryo transfer.

  FIG. 12 shows a comparison between the laparoscopic embryo transfer method and the transvaginal embryo transfer method. Transvaginal transplantation can be performed in a shorter time than open embryo transfer. In addition, wound adhesion occurs in the open embryo transfer method, whereas adhesion does not occur in the transvaginal embryo transfer method. In addition, it is necessary to perform various post-operative care in the laparoscopic embryo transfer method, but in the transvaginal embryo transfer method, it is sufficient to administer antibiotics. Furthermore, in the case of a common marmoset, if the transplantation is not successful, the open embryo transfer method requires 2 months to perform the next embryo transfer using the individual, but the transvaginal embryo transfer method requires 3 weeks. Later, embryo transfer can be performed again. As described above, the transvaginal embryo transfer method using the egg collection and embryo transfer device of the present invention makes it possible to make the embryo transfer technique non-invasive, and the interval of embryo transfer per experimental animal is shortened. The number of animals to be played can be reduced.

Example 2 Fertilized Egg Transplantation Using the Egg Collection and Embryo Transfer Device of the Present Invention Fertilized eggs were collected by a uterine perfusion method in which the uterus was perfused using a female marmoset as a donor of the fertilized egg. At this time, the transvaginal egg collecting method for performing surgical operation as a control and the transcutaneous egg for collecting the needle by inserting a needle from the skin together with the transvaginal egg collecting method using the egg collecting and embryo transfer device of the present invention The sampling method was performed. Egg collection was performed 52 times by each method, and egg collection efficiency was examined.

  FIG. 13B shows the number of times eggs can be collected by each method (number of times of collection), the number of eggs collected, the number of modified eggs, the collection rate, and the average number of eggs collected. FIG. 13A is a graph showing the average number of eggs collected. As shown in FIGS. 13A and B, the average number of eggs collected by the transvaginal egg collection method using the egg collection and embryo transfer device of the present invention is the same as the results obtained by the laparotomy egg collection method or the percutaneous egg collection method, or these It was better than the conventional method.

FIG. 14 shows a comparison of the laparotomy egg collection method, the transdermal egg collection method, and the transvaginal egg collection method of the present invention.
Eggs could be collected with the same efficiency as the conventional transdermal egg collection method. The transvaginal egg collection method is superior in that it is a non-invasive method compared to the laparotomy egg collection method and the transdermal egg collection method. As the technician becomes skilled, eggs can be collected more efficiently.

  The egg collection and embryo transfer device of the present invention can be used for fertilized egg collection and embryo transfer of laboratory animals such as small monkeys, and genetic modification, production of chimeric animals and artificial propagation can be performed using the device. Can do.

DESCRIPTION OF SYMBOLS 1 Inner core 2 Inner core needle part 3 Inner core holding part 4 Catheter 5 Catheter tube part 6 Catheter connection part 7 Guide catheter 8 Guide catheter tube part 9 Guide catheter connection part 10 Guide tube 11 Uterine ostium 12 Vaginal opening 13 Uterine cavity 14 Vagina 15 Fertilized egg 16 Embryo

Claims (3)

An inner core made of a metal needle; a catheter composed of a flexible tube made of a flexible material with a distal end and a proximal end opened; and made of a flexible material into which the inner core or catheter can be slidably inserted. A rigid material having a cage elasticity, having a distal end and a proximal end opened, and a distal end having a tapered shape; and a rigid material into which the guide catheter or a combination of the guide catheter and the inner core or the catheter can be inserted. It includes a guide tube that is made in a component is inserted into the uterus in a state of inserting the inner core to the guide catheter, used to insert a vent away catheter subsequent inner core, the inner core and the catheter from the guide catheter even longer, the guide catheter is longer than the guide tube, in a state in which the catheter is inserted into the guide catheter, uterine from the tip of the catheter An egg in the perfusate that flows into the guide catheter from the opening at the distal end of the guide catheter and flows out of the body from the opening at the proximal end of the guide catheter can be collected. , The inner core needle size is 24-26 gauge and length is 100-200mm, catheter size is 22-24 gauge, tube part length is 100-200mm, guide catheter size is 18-20 a length 70~130mm of are tube portions gauge the size of the guide pipe is that there length with an outer diameter 3 to 10 mm 50 to 100 mm, small monkeys for eggs adopted take-up device. Further comprising a syringe, according to claim 1 Symbol placement egg adopted take-up device. A method of collecting fertilized eggs from the uterus of small monkeys using egg adopted take-up device according to claim 1 or 2,
Inserting a guide tube into the vagina from the vaginal opening of a small monkey;
An inner core is inserted into the guide catheter to form a double-structure catheter, and the double-structure catheter is inserted through the guide tube until the inner core and the distal end of the guide catheter reach the uterus;
After insertion, removing the inner core from the guide catheter;
Inserting a catheter into the guide catheter;
Flowing perfusate into the uterus through an opening at the proximal end of the catheter;
With the catheter inserted into the guide catheter, perfusate that enters the guide catheter through the opening at the distal end of the guide catheter, passes through the guide catheter and flows out of the body from the opening at the proximal end of the guide catheter. A method for collecting fertilized eggs from the uterus of a small monkey, which comprises collecting and collecting the fertilized eggs contained in the perfusate.

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