Rabbit embryo transfer technique

Summary

Embryo transfer, commonly known as artificial insemination, is a technique in which the early embryo of a female animal, or an embryo obtained by in vitro fertilization or other means, is transplanted into a surrogate female animal of the same species and physiological state, so that the embryo can continue to develop into a new individual. Rabbits, as experimental animals, are superior to other animals, and can be used as the basic research for embryo transfer in all livestock, and can provide embryo sources for new techniques of embryo manipulation related to genetic engineering of livestock (embryo segmentation, nuclear transfer, transgenesis, etc.).

Principle

The principle of embryo transfer is based on several physiological foundations:

① The physiological changes in the reproductive organs of the donor and recipient of the same species of animal are the same, providing the same physiological environment for the transfer of the donor embryo into the recipient.

② Early embryos are free for a certain period of time, providing the possibility of embryo collection.

(iii) Recipients basically do not undergo immune rejection of foreign embryos transferred into the uterus, providing the possibility of embryo survival in the recipient.

(iv) The donor embryo can establish normal physiological and organizational links with the recipient uterus, but the genetic characteristics of the donor embryo are not affected in any way during conception.

Operation method

Rabbit embryo transfer technique

Principle

The principle of embryo transfer is based on the following physiological foundations: ① The physiological changes in the reproductive organs of the donor and recipient of the same species of animal are the same, providing the same physiological environment for the transfer of the donor embryo into the recipient. ② Early embryos are in a free state for a certain period of time, providing the possibility of embryo collection. (iii) Recipients basically do not undergo immune rejection of foreign embryos transferred into the uterus, providing the possibility of embryo survival in the recipient. (iv) The donor embryo can establish normal physiological and organizational links with the recipient uterus, but the genetic characteristics of the donor embryo are not affected in any way during conception.

Materials and Instruments

Equipment:
① syringe
② Special pipette with an inner diameter of 0.5 to 1 mm
③An environmentally clean animal room with light conditions of 8:16 (light:dark) hours.
Reagents:
①Materials:
②GnRH hormone
③Egg transfer solution

Move

The basic process of embryo transfer in rabbits can be divided into the following steps:

1. Recipient rabbits are treated with simultaneous estrus and pseudo-pregnancy, and each recipient rabbit is injected intramuscularly with 15 μg of GnRH hormone (the estrus period is 22 hours later than that of the embryo donor), and then caged together with the ligated male rabbits.

2. Embryos of all periods were transferred into the oviducts. First, the trumpet was identified, and then a special pipette with an inner diameter of 0.5-1 mm was sucked up the embryos to be transferred together with 0.05 ml of egg transfer solution and carefully inserted into the trumpet, slowly placed into the oviduct, and then the embryos were transferred.

3. Both oviducts were transferred, with 3~5 embryos on each side.

4. After embryo transfer, the recipient rabbits were kept in an environmentally clean-grade animal rearing room with a light condition of 8:16 (light:dark) hours.

5. Pregnancy was checked by abdominal palpation 14-16 days after transfer. The pregnancy expired on day 31-32 of the recipient's gestation and live pups were obtained by natural delivery (day 30-31) or cesarean section (day 32).

Caveat

1. Embryo transfer should be carried out under aseptic conditions as far as possible, the procedure should be accurate and rapid, the shorter the time the embryo stays outside the room, the better, the room temperature should be at 18-25 ℃ during the embryo transfer, and the whole procedure should take about 30 minutes.

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