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A culture medium for promoting immature oocyte maturation in vitro and its application

An oocyte, in vitro maturation technology, applied in the field of reproductive biology, can solve the problems of in vitro maturation and cultivation of uncertain tertiary cumulus-oocyte complexes, etc. The effect of improving embryo quality

Active Publication Date: 2020-07-21
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is still uncertain whether BAPTA-AM can promote the in vitro maturation of the tertiary cumulus-oocyte complex and be used for embryo production, and there is no research report in this regard

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The immature oocyte maturation culture in vitro of embodiment 1 bovine

[0033] (1) Collection of oocytes

[0034] Collect the ovaries of dairy cows from the slaughterhouse, put them in a thermos flask containing double-antibody (0.024g / L penicillin + 0.02g / L streptomycin), keep the temperature at 35°C-37°C, and send them within 6 hours Back to the lab. The ovaries were washed 2 to 3 times with normal saline containing double antibodies, and then washed with 2ml egg extraction solution (9.5g / L M199+10%FBS+0.042%NaHCO3+0.2383%Hepes+0.2603%Hepes-Na+0.001%heparin sodium + 0.005% penicillin + 0.0065% streptomycin) into a 10 ml syringe with a 18-gauge needle to extract the oocyte-follicular fluid complex of 3-8 mm follicles. Put the oocyte extraction liquid after absorbing the oocytes into a domestic petri dish with a diameter of 9 cm, and select first-class COCs (cumulus-oocytes with uniform cytoplasm and 5 or more layers of surrounding granulosa cells) under a stereo mic...

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PUM

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Abstract

The invention relates to an immature oocyte in vitro maturation promoting culture medium and application thereof. The immature oocyte in vitro maturation promoting culture medium is screened out through research on effects of different additives on immature oocytes and prepared by adding 20-100 mu M of BAPTA-AM in normal oocyte in vitro maturation culture solution. The immature oocyte in vitro maturation promoting culture medium is safe and free to side and toxic effects, enables meiosis of the immature oocytes, particularly those in level 3 cumulus-oocyte complexes, improves the in vitro development of the oocytes, promotes maturation, improves the cleavage rate and the blastula development rate after the mature oocytes are parthenogenetically-activated and further improves embryo quality. Application of the immature oocyte in vitro maturation promoting culture medium provides favorable support for the technology of in vitro propagation of animal embryos.

Description

technical field [0001] The invention relates to a culture medium for promoting in vitro maturation of immature oocytes and application thereof, belonging to the technical field of reproductive biology. Background technique [0002] In terms of reproductive biology, embryo production requires a large number of egg sources. Generally, oocytes obtained directly from the ovary of slaughtered animals are used, or oocytes are obtained from the ovaries of living animals by live egg collection. However, due to the The obtained oocytes are not fully mature, so a period of in vitro maturation and culture is required. After the immature oocytes are matured, the subsequent stages of in vitro fertilization and development into embryos can be completed, and reproduction and production can be truly realized. [0003] Mammalian ovaries contain a large number of immature oocytes. Therefore, taking immature oocytes directly from the ovary for in vitro maturation and culture is the most effect...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2500/36
Inventor 侯云鹏胡洪梅傅祥伟莫显红
Owner CHINA AGRI UNIV
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