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Method for promoting sheep in-vitro embryo development by using follistatin

A technology for embryo development and follistatin, which is applied in the field of bioengineering to achieve the effect of improving the rate of developing blastocysts

Inactive Publication Date: 2015-06-10
新疆畜牧科学院生物技术研究所
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Problems solved by technology

But at present, there is no report on the effect of Follistatin on the development of sheep embryo culture in vitro

Method used

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Examples

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Embodiment 1

[0012] Example 1: Extract the sheep ovary within 30 minutes of slaughter, put it into the physiological saline containing 1000IU / ml penicillin and 1000IU / ml streptomycin at a temperature of 30-35°C, and wash it with physiological saline for 3-4 hours within 3 hours. 2 times: Use a 10ml syringe with 2ml of ovum extraction fluid and a No. 9 needle to aspirate follicles with a size of 2-5mm on the surface of the ovary, put the recovered follicles in the syringe into a 35-60mm dish, and pick them out under a microscope Oocytes, washed 3 times with in vitro maturation solution; in the condition of 5% CO 2 , 95% air, CO at a temperature of 38.6°C 2 Cultivate in the incubator for 22-24 hours, wash 2-4 times with in vitro fertilization fluid; add 25-30 pieces / drop into 50-70 μl in vitro fertilization droplets that were balanced 2 hours ago; thaw the semen in a water bath, Transfer to the bottom of the test tube containing in vitro fertilization fluid that was equilibrated 2 hours ago...

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Abstract

The invention belongs to the technical field of bioengineering, and relates to a method for promoting sheep in-vitro embryo development by using follistatin, which comprises the following steps: (1) acquiring a slaughtered sheep ovary; (2) extracting follicles for later use; (3) culturing selected oocytes in a CO2 incubator, and adding into in-vitro fertilization solution drops; (4) adding the in-vitro fertilization drops, and carrying out co-incubation with the oocytes; (5) taking out the early embryo, transferring into a 10 mu g / L follistatin in-vitro culture solution, washing 3-4 times, and culturing for 24-26 hours; and (6) taking out the embryo, washing with the in-vitro culture solution, calculating the merogenesis rate, and calculating the blastula rate after culturing for 144-168 hours. By using the follistatin to treat the early embryo subjected to sheep oocyte in-vitro fertilization, the method can obviously enhance the in-vitro embryo development blastula rate.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for promoting embryonic development in vitro of sheep by utilizing follistatin. Background technique [0002] Follistatin is a cysteine-rich glycosylated single-chain polypeptide isolated from bovine and pig follicular fluid by Robertson and Ueno in 1987. It can inhibit the secretion of follicle-stimulating hormone by combining with activin protein. Follistatin can not only hinder the function of activin protein, but also reduce the functions of other TGF-β family members including myostatin (MSTN), GDF-9, bone morphogenetic proteins (BMPs). By binding to these TGF-β family members, follistatin is widely involved in the regulation of multiple physiological functions of organisms, including reproduction, growth and development, neural induction, osmotic pressure regulation, myogenesis and immune regulation. But at present, there is no report on the ef...

Claims

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Application Information

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IPC IPC(8): C12N5/073
Inventor 黄俊成汪立芹吴阳升林嘉鹏何宗霖唐森阿米娜·阿布都瓦力
Owner 新疆畜牧科学院生物技术研究所
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