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A kind of cryopreservation solution and cryopreservation method for oocytes in gv stage

A technology for cryopreservation of oocytes, applied to germ cells, animal cells, vertebrate cells, etc., can solve the problems of oocyte fertilization cleavage rate and low blastocyst development rate, so as to improve developmental ability and reduce damage , low-cost effect

Inactive Publication Date: 2018-03-06
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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AI Technical Summary

Problems solved by technology

[0006] In order to solve the problem of low oocyte fertilization cleavage rate and blastocyst development rate after oocyte cryopreservation and thawing, the present invention provides a GV stage oocyte cryopreservation solution and a cryopreservation method

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  • A kind of cryopreservation solution and cryopreservation method for oocytes in gv stage

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specific Embodiment approach 1

[0024] Specific embodiment 1: In this embodiment, oocyte cryopreservation equilibrium solution is made up of conventional culture medium, fetal bovine serum, vitamin B 12 , folic acid, ethylene glycol, DMSO and trehalose; the volume concentration of fetal bovine serum in the balance solution is 10%, vitamin B 12 The concentration of folic acid is 2mg / ml, the concentration of folic acid is 100μM, the volume concentration of ethylene glycol is 7.5%, the volume concentration of dimethyl sulfoxide is 7.5%, and the concentration of trehalose is 0.25M; the conventional culture medium is DMEM culture medium , TCM199 culture medium, M16 or MEM culture medium.

specific Embodiment approach 2

[0025] Specific embodiment two: present embodiment oocyte cryopreservation liquid is made of conventional culture medium, fetal bovine serum, vitamin B 12 , folic acid, ethylene glycol and DMSO; the volume concentration of fetal bovine serum is 10%, vitamin B 12 The concentration of folic acid is 2mg / ml, the concentration of folic acid is 100μM, the volume concentration of ethylene glycol is 15%, and the volume concentration of DMSO is 15%. The conventional culture medium is DMEM medium, TCM199 medium, M16 or MEM medium.

specific Embodiment approach 3

[0026] Specific embodiment three: In this embodiment, oocyte cryopreservation thawing liquid is divided into two kinds of thawing liquid I and thawing liquid II; 12 , folic acid and trehalose, the volume concentration of fetal bovine serum in the thawing solution is 10%, vitamin B 12 The concentration of trehalose is 2mg / ml, the concentration of folic acid is 100μM, the concentration of trehalose in thawing solution Ⅰ is 1M, and the concentration of trehalose in thawing solution Ⅱ is 0.5M; among them, the conventional culture medium is DMEM culture medium, TCM199 culture medium, M16 or MEM medium.

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Abstract

A cryopreservation solution and cryopreservation method for oocytes in the GV stage. It relates to an oocyte cryopreservation solution and a cryopreservation method. The invention solves the problem of low survival rate and development rate after cryopreservation and thawing of the existing oocytes. Oocyte freezing uses cryoloop as carrier, conventional culture medium as substrate, DMSO and ethylene glycol as cryoprotectants, and vitamin B12 and folic acid are added during freezing and thawing. The invention can effectively improve the oocyte survival rate and in vitro development rate after thawing.

Description

technical field [0001] The invention relates to an oocyte cryopreservation solution and a cryopreservation method. Background technique [0002] The cryopreservation of mammalian oocytes refers to the suppression of all metabolic activities in the cells under ultra-low temperature (-196°C) conditions, and the cells are preserved for a long time. After thawing in a certain way, their developmental potentials such as fertilization and cleavage can be restored. a preservation technique. Oocyte cryopreservation can make full use of oocytes from various sources, and can provide a large number of experimental materials for the research and application of embryo engineering technologies such as in vitro fertilization, nuclear transfer, and transgenesis, and is not limited by time and space. [0003] The cryopreservation technology of oocytes comes from cryopreservation of embryos. Due to a series of special properties of the cells themselves, cryopreservation is more difficult, an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/075
Inventor 曹新燕李井春徐超赵家平赵伟刚李雁冰张宇飞赵蒙薛海龙刑明杰王丽英
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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