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Cow ovum cell in vitro ripening culturing liquid containing tea polyphenol and its culturing method

A technology of in vitro maturation culture and oocyte, applied in the field of livestock embryo engineering, which can solve the problems of excessive free radicals and low early embryo development rate.

Inactive Publication Date: 2008-11-12
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to solve the problem of too many free radicals in the in vitro culture medium of oocytes in the prior art, which leads to a low early embryonic development rate after oocyte fertilization, and provides a method for timely removing oocytes and oocytes. Tea polyphenol-containing bovine oocyte in vitro maturation medium and culture method for excessive free radicals in culture environment and function of improving embryo development rate after in vitro fertilization of oocytes

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Preparation of oocyte maturation medium in vitro containing tea polyphenols

[0022] Accurately weigh 4 milligrams of tea polyphenols, dissolve them with 20 milliliters of M199 basal culture medium (purchased from GI BCO, USA), and then add 10 milliliters of calf serum and 10,000 international units of penicillin to the culture medium. Streptomycin, stirred gently, then added M199 basal culture medium to a volume of 100 ml, and stirred again. After filtration and disinfection with 0.22 μM microfiltration membrane (Millipore Corporation, Bedford.Mass.USA), store at 4°C until use.

Embodiment 2

[0023] Example 2: Preparation of oocyte maturation medium in vitro containing tea polyphenols

[0024] Accurately weigh 3 milligrams of tea polyphenols, dissolve them with 20 milliliters of DMEM basal culture fluid (purchased from G I BCO, U.S.), then add 10 milliliters of fetal bovine serum and 10,000 international units of penicillin to the culture fluid, 10,000 micrograms Streptomycin, stir gently, add DMEM basal culture medium to 100 ml, stir and mix. After filtration and disinfection with a 0.22 μM microfiltration membrane (Millipore Corporation, Bedford. Mass. USA), store at 4° C. for use.

Embodiment 3

[0025] Example 3: Tea polyphenol-containing culture fluid is used for in vitro maturation of dairy cow oocytes

[0026] Dairy cow oocytes were matured and cultured in the control culture solution without tea polyphenols and in vitro maturation culture solution containing tea polyphenols prepared in Example 1, and then fertilized in vitro. Early embryos after fertilization continue to be cultured for 8 days. On the second day of cultivation, the cleavage rate of oocytes in the control medium without tea polyphenols after fertilization was 71.3%, and the oocytes in the in vitro maturation medium containing tea polyphenols prepared in Example 1 The division rate after fertilization was 79.5%, which was 8.2% higher than that of the control group. On the 8th day of cultivation, the blastocyst stage development rate after fertilization of oocytes in the control culture medium not containing tea polyphenols was 19.3%, and in the in vitro maturation culture medium containing tea poly...

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Abstract

The present invention provides an in vitro mature culture fluid containing tea-polyphenol for bovine ovocyte and its culture method. The described in vitro mature culture flud uses conventional culture fluid as matrix, in the described matrix 20-150 microgram / mL of tea-polyphenol, 100 IU / mL of penicillin, 100 microgram / mL of streptomycin and serum whose volume percentape content is 8%-20% are also contained. Said invention can make tea-polyphenol be used in the field of bovine ovocyte in vitro mature culture so as to raise early embryonic in vitro development rate after the bovine ovocyte in vitro fertilization.

Description

(1) Technical field [0001] The invention relates to a bovine oocyte in vitro maturation culture solution containing tea polyphenols and a culture method thereof, belonging to the technical field of livestock embryo engineering. (2) Background technology [0002] With the development of modern reproductive biotechnology and the gradual improvement of embryo in vitro production technology, the use of in vitro fertilization technology to produce in vitro embryos presents a potential broad application prospect. In vitro production of livestock embryos, especially somatic cell nuclear transfer, is an effective way to rapidly expand the number of excellent populations. A sufficient number of mature oocytes is a prerequisite for in vitro production of embryos. Extracting oocytes with a syringe from ovaries obtained from slaughterhouses and culturing them in vitro is the most effective and economical way to obtain mature oocytes. Since the concentration of oxygen in the reproducti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/06C12N5/075
Inventor 俞颂东王争光陈阿琴
Owner ZHEJIANG UNIV
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