Rabbit chimera technology

Summary

A mammalian chimera is an individual that develops from a polymerized embryo composed of two or more cells with different heritabilities. Cells of different origins within a chimeric embryo regulate and interact with each other to the extent that they work together to complete embryonic development. Because chimeras can carry appropriate genetic markers and express them in individual development, they have become the most favorable experimental material and genetic research model for studying mammalian individual development.

Operation method

Rabbit chimera technology

Materials and Instruments

Equipment:
① syringe
② glass microneedle
③ light conditions for 8: 16 (light: dark) hours of the environment of clean-grade animal rearing room
Reagents:
①Material: rabbit
② Protease
③ 0.1% EDTA without Ca, Mg
2
Hanks solution
④hCG

Move

The basic process of the rabbit chimera technique can be divided into the following steps:

1. The donor embryo is obtained from the uterus, and the mucin layer and zona pellucida of the washed-out embryo are removed by protease.

2. Using a homemade glass microneedle, the embryo is cut into two parts, one half containing ICM and a small number of trophoblast cells, and one half containing only trophoblast cells.

3. The ICM-containing half embryo was placed in Ca and _{Mg2Hanks-free} solution containing 0.25% trypsin and 0.1% EDTA, and gently blown with a microfuge tube for several times to obtain the isolated ICM cells.

4. ICM cells at 96 hours and 120 hours after mating were injected into the blastocoel cavity of 96-hour recipient embryos, with 10-15 cells injected into each recipient embryo.

5. A natural estrous female rabbit synchronized with the recipient embryo was selected and injected intravenously with 50 U of hCG as a pseudopregnant rabbit.

6. The injected blastocysts were transferred to the uterus of the sham pregnant rabbits.

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