Embryo Transfer Into Rabbit: A Procedure for Laparoscopy-Guided Transfer of Embryos Into Oviduct of Ovulating Recipient Female Rabbit Model
Transcription
On the day of the transfer, rinse the fresh or thawed embryos in 37 degrees Celsius base medium under a stereo microscope, and attach an appropriately configured 17 gauge epidural catheter to a 1-milliliter syringe. Aspirate one centimeter of base medium into the catheter followed by a small air bubble. Then, aspirate 5 to 7 embryos and 10 microliters of base medium, followed by another small air bubble in a final one centimeter of base medium.
Once the embryos have been loaded, place an anesthetized recipient rabbit in a cage on a warming stage, and apply ointment to the animal's eyes. After confirming a lack of response to pedal reflex, shave the fur from the ventral abdomen, clean the surgical area with chlorhexidine gluconate soap, and remove any remaining hair.
Cover the area with a sterile drape with a hole for the surgical area, and insert one endoscopic trocar 5 centimeters into the abdominal cavity, 2 centimeters caudal to the xiphoid process, and use a pressure-regulating mechanical insufflator to inflate the peritoneal cavity.
Insert the endoscope camera through the endoscopic trocar, and insert the 17-gauge epidural needle into the inguinal region 2 to 3 centimeters from the infundibulum. Insert the loaded catheter through the epidural needle into the abdomen, and insert 1 to 2 centimeters of the epidural catheter through the infundibulum in the ampulla.
Gently depress the catheter plunger to release the embryos into the oviduct. Both air bubbles must exit from the catheter. Remove the catheter just after the embryos have been released, and aspirate and release the remaining medium to confirm the absence of the embryos.
After confirming a successful transfer of the embryos, remove the epidural needle and the endoscope camera, and release the abdominal carbon dioxide through the endoscopic trocar. Clean the trocar incision with chlorhexidine solution, and close the wound with a micronized aluminum and a plastic dressing. Then, treat the animal with the appropriate antibiotics and analgesics and monitor the animal until full recovery.
