Vitrification freezing and thawing method for embryos

Abstract

The invention discloses a vitrification freezing and thawing method for embryos, and belongs to the field of embryo biotechnology. The technical scheme comprises: a freezing liquid and a thawing liquid are put into thin tubes in a segmented manner, each section of the liquids is separated by air, embryos are placed in one section of the freezing liquids, and then the thin tubes are sealed and putinto liquid nitrogen. The characteristics are that: two sections of the freezing liquids and two sections of the thawing liquids are respectively put into the thin tubes, the volume ratio of the freezing liquid to the thawing liquid is less than or equal to 1:5, or the concentration of an anti-freezing protectant in the mixed liquid after mixing is within 2.0 Mol / L. A simple thawing and transplantation method of the invention is that: the thin tubes used for cryopreservation of embryos are taken out from liquid nitrogen at room temperature, the thin tubes are subjected to be in air bath for 4-6 s and water bath at 20-25 DEG C, the thin tubes are taken out, the thin tubes are reversed back and forth to make the freezing liquid mix evenly, and embryos are transferred. The method greatly simplifies a thawing procedure of embryos after vitrification freezing, and detoxification inside tubes and direct transplantation of embryos are realized. The method is simple and convenient, and high inoperability, and thus the method is beneficial to popularization and application.

Description

technical field [0001] The invention relates to a vitrified embryo freezing, thawing and direct transplantation method, in particular to a mammalian embryo freezing, thawing and transplantation method, which belongs to the field of embryonic biotechnology Background technique [0002] In 1972, Whittingham first used dimethyl sulfoxide (Dimethylsulfoxide: DMSO) as the main anti-cryoprotectant to cryopreserve 2-8-cell embryos of mice successfully, and transplanted them to give birth to offspring. The concentration of the freezing solution they adopted was 1.5Mol / L, and a programmed cooling freezer was used. After freezing and thawing, the embryo survival rate was 50%-70%, the embryo pregnancy rate was 65%, and the litter rate was 48%. Since the invention of the programmed freezing method, researchers from various countries have repeatedly tested and screened various cryoprotectants, so that the survival rate of embryos after freezing and thawing can reach more than 90%, while...

AI Technical Summary

Problems solved by technology

The thawing process is not only very cumbersome, but also easily loses embryos, requiring skilled technicians

These conditions have restricted the popularization and application of vitrification freezing method in production.

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