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Ox embryo in vitro culturing liquid containing tea polyphenol and its culturing method

An in vitro culture, bovine embryo technology, applied in embryonic cells, tissue culture, biochemical equipment and methods, etc., can solve the problems of free radicals and low early embryo development rate.

Inactive Publication Date: 2008-09-17
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to solve the problem of too many free radicals in the in vitro culture solution of bovine embryos in the prior art, resulting in a low embryonic early development rate, and provides a method for timely removing excessive free radicals in the embryonic cells and in the culture environment, improving the Tea polyphenol-containing bovine embryo in vitro culture solution and its culture method for early embryo in vitro development rate function

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Preparation of tea polyphenol-containing bovine embryo in vitro culture fluid

[0020] Accurately weigh 5 mg of tea polyphenols, dissolve them in 20 mL of SOF basal culture medium, then add 800 mg of bovine serum albumin, 10,000 international units of penicillin, and 10,000 micrograms of streptomycin to the culture medium, stir evenly, and then Add SOF basal culture medium to 100 ml, and stir again. After filtration and disinfection with a 0.22 μM microfiltration membrane (Millipore Corporation, Bedford. Mass. USA), store at 4° C. for use.

Embodiment 2

[0021] Embodiment 2: The nutrient solution containing tea polyphenols is used for in vitro culture of dairy cow embryos

[0022] Cow oocytes were matured in M199 medium (purchased from GIBCO, USA) containing 100 IU / mL penicillin, 100 μg / mL streptomycin, and 8% to 20% serum by volume, and then fertilized in vitro . The early embryos after fertilization were further cultured for 8 days in the SOF basal culture medium without tea polyphenols (control group) and the in vitro culture medium of bovine embryos containing tea polyphenols prepared in Example 1, respectively. On the second day of culturing, the early embryo division rate in the control group was 69.1%, and the early embryo division rate in the tea polyphenol-containing bovine embryo in vitro culture medium prepared in Example 1 was 78.7%, which was 9.6% higher than that of the control group . On the 8th day of culturing, the blastocyst stage development rate of early embryos in the control group was 19.1%, and the bla...

Embodiment 3

[0023] Example 3: The tea polyphenol-containing culture solution is used for the in vitro culture of cattle embryos

[0024] Cattle oocytes are matured and cultured in M199 medium containing 100 IU / mL penicillin, 100 μg / mL streptomycin and 8%-20% serum by volume, and then fertilized in vitro. The early embryos after fertilization were further cultured for 8 days in the SOF basal culture medium without tea polyphenols (control group) and the in vitro culture medium of bovine embryos containing tea polyphenols prepared in Example 1, respectively. On the second day of cultivation, the early embryo division rate in the control group was 72.4%, and the early embryo division rate in the tea polyphenol-containing bovine embryo in vitro culture medium prepared in Example 1 was 81.3%, which was 8.9% higher than that of the control group . On the 8th day of culturing, the blastocyst stage development rate of early embryos in the control group was 21.7%, and the blastocyst stage develop...

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PUM

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Abstract

The present invention provides an in vitro culture fluid containing tea-polyphenol for bovine embryo. The described bovine embryo in vitro culture fluid uses conventional culture fluid as matrix. In the described matrix 20-80 microgram / mL of tea-polyphenol, 100IU / mL of penicillin, 100 microgram / mL of streptomycin and 5-10 mg / mL of serum albumin are also contained. Said invention makes the tea-polyphenol be used in the field of bovine embryo in vitro culture so as to raise the early in vitro development rate of bovine in vitro fertilized embryo.

Description

(1) Technical field [0001] The invention relates to a bovine embryo in vitro 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. Although live offspring have been obtained through in vitro fertilization technology, the in vitro development rate of early embryos after in vitro fertilization is low, which limits the popularization and application of embryo in vitro production in practice. [0003] Since the concentration of oxygen in the reproductive tra...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/06A61D19/00C12N5/073
Inventor 王争光俞颂东陈阿琴
Owner ZHEJIANG UNIV
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