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Method for improving quality of external fertilization embryo of oocyte of sheep

A technology of oocytes and in vitro fertilization, applied in the direction of artificial cell constructs, germ cells, animal cells, etc., can solve the problems of few, improve the quality of sheep oocytes in vitro fertilized embryos, and the developmental ability of embryos has not been significantly improved.

Inactive Publication Date: 2013-11-27
新疆维吾尔自治区畜牧科学院中国-澳大利亚绵羊育种研究中心
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Problems solved by technology

③Zygote, 1 (2011), 1-5, author: C.S.Oliveira et al. published "The Effect of Histone Deacetylase Inhibitor TSA in Bovine Embryo Culture in Vitro", and concluded that TSA has a significant effect on the development of bovine 8-cell embryos. No significant improvement in ability
The above-mentioned existing methods are used in somatic cell nuclear transfer and in vitro fertilized embryo culture in cattle, pigs, and mice, but there are few reports on improving the quality of sheep oocytes in vitro fertilized embryos

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  • Method for improving quality of external fertilization embryo of oocyte of sheep

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Embodiment

[0025] 1) Remove sheep ovaries that were slaughtered within 30 minutes, put them in normal saline containing 1000IU / ml penicillin and 1000IU / ml streptomycin at a temperature of 32°C, wash them with normal saline three times within 3 hours; A 10ml syringe with a No. 9 needle is used to suck follicles 2-8mm in size from the surface of the ovary, and the follicle fluid recovered from the syringe is placed in a 45mm dish, and the oocytes are picked out under a microscope, and washed with the egg extraction solution for 3 times, then washed 3 times with mature solution, placed in CO 2 Cultivate in the box for 24h, CO 2 The condition of the box is 5% CO 2 , 95% air, temperature 38.6°C; at the end of the cultivation, wash with TSA-containing in vitro fertilization solution for 3 times, and add 30 pieces / drop into 70 μl of TSA-containing in vitro fertilization droplets that were balanced 2 hours ago;

[0026] 2) Thaw the semen in a water bath, transfer it into a test tube of in vitr...

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Abstract

The invention provides a method for improving quality of an external fertilization embryo of an oocyte of a sheep. The method relates to TrichostatinA (TSA) which is used for treating the oocyte of the sheep and can greatly improve the quality of the external fertilization embryo of the oocyte of the sheep, by adopting the method improving the quality of the external fertilization embryo of the oocyte of the sheep through TSA, after 168 hours, the blastocyst rate and the number of the obtained blastomeres are remarkably higher than those of the matched group, therefore, the method has better scientificity and practicability.

Description

technical field [0001] The invention relates to a method for treating sheep oocytes with Trichostatin A (TSA), which can significantly improve the quality of sheep oocytes in vitro fertilized embryos. Background technique [0002] Trichostatin A (TSA) is a histone deacetylase (Histone deacetylase, HDACs) inhibitor. DNA and histones are the main components of chromatin. Chromatin structure is closely related to gene activity, and chromatin structure is modified through histone acetylation and deacetylation, which plays an important role in DNA replication, gene transcription, and cell cycle regulation. At present, TSA is mainly used in the research of improving the developmental ability of cloned embryos. Its application in mammalian embryonic development in vitro has only been reported in cattle and mice, and there is no report on its application in sheep oocytes. At the same time, the number of blastocyst cells also reflects the quality of embryos to a certain extent. ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075C12N5/071
Inventor 黄俊成汪立芹杨楠林嘉鹏吴阳升唐森赵云程
Owner 新疆维吾尔自治区畜牧科学院中国-澳大利亚绵羊育种研究中心
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