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Vitrification freezing, simple defreezing and direct implanting method for embryo

A vitrification freezing and transplantation technique, applied in the field of embryo biology, can solve the problems of easy loss of embryos, cumbersome thawing process, restricting the popularization and application of vitrification freezing method, etc., and achieves the effect of good effect, strong operability and length increase.

Inactive Publication Date: 2005-08-17
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The thawing process is not only very cumbersome, but also easy to lose embryos, requiring skilled technicians
These conditions have restricted the popularization and application of vitrification freezing method in production.

Method used

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  • Vitrification freezing, simple defreezing and direct implanting method for embryo

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Embodiment Construction

[0019] in as figure 1 Among the shown packing methods, 1 is a cotton plug and sealing; 2 is a sealing liquid (its composition is the same as that of 6); 3 is an air section; 4 is a freezing liquid containing embryos; 5 is a freezing liquid; 6 is a thawing liquid.

[0020] The inventive point of the vitrified embryo freezing method of the present invention is that the configuration of the freezing liquid and the thawing liquid in the thin tube is different from other methods. Depend on figure 1 It can be seen that, compared with the "5-section method" of the existing vitrified embryo freezing technology, this tube arrangement reduces the number of air sections and shortens the length of the freezing liquid section, which can ensure that more thawing liquid is used. To dilute the high-concentration freezing solution to complete the process of removing the antifreeze protection agent inside the embryo. Because the freezing liquid section is very short, if the embryos are loaded...

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Abstract

The present invention discloses embryo vitrifying freezing, defreezing and directly transplanting method, and belongs to the field of embryo biotechnology. The technological scheme includes setting freezing liquid and defreezing liquid in different sections of fine tube separated with air, setting embryo in one section of the freezing liquid, sealing ends of the fine tube, and throwing the fine tube into liquid nitrogen; and features that there are two sections of freezing liquid and two sections of defreezing liquid with volume ratio between the freezing liquid and defreezing liquid of 1 to 5, or antifreeze protecting agent concentration in the mixed liquid of 2.0 mol / L. The simple defreezing and transplanting process includes taking out the fine tube from liquid nitrogen, air bathing for 4-6 sec, water bathing at 20-25 deg.c, shaking for homogeneous mixing of freezing liquid and transplanting. The present invention realizes the simple tube detoxication of embryo and direct transplanting.

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, and belongs to the field of embryonic biotechnology. technical background [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 used was 1.5Mol / L, and a programmed cooling freezer was used. After freezing and thawing, the embryo survival rate was 50% to 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/08
Inventor 朱士恩杨中强侯云鹏
Owner CHINA AGRI UNIV
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