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In vitro maturation medium of immature oocytes and its application

A technology for in vitro maturation culture and oocyte, applied in cell culture medium, cell culture active agent, artificial cell construct, etc. rate effect

Active Publication Date: 2021-03-02
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unfortunately, the number of non-human embryos produced in vitro is significantly lower than in vivo, which reflects the suboptimal oocyte culture system

Method used

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  • In vitro maturation medium of immature oocytes and its application
  • In vitro maturation medium of immature oocytes and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] The preparation of embodiment 1 in vitro maturation culture medium

[0075] Dissolve FBS, HDACi 4b, CNP, glutamine, penicillin and streptomycin in TCM-199 to prepare a solution containing 10% FBS, 5μM HDACi 4b, 60ng / mL CNP, 0.1mg / mL glutamine, 100IU / mL of penicillin and 100IU / mL of streptomycin in TCM-199 medium is the in vitro maturation medium.

Embodiment 2

[0076] The preparation of embodiment 2 in vitro maturation culture medium

[0077] Dissolve FBS, HDACi 4b, CNP, glutamine, penicillin and streptomycin in TCM-199 to prepare a solution containing 5% FBS, 3μM HDACi 4b, 80ng / mL CNP, 0.05mg / mL glutamine, 100IU / mL of penicillin and 100IU / mL of streptomycin in TCM-199 medium is the in vitro maturation medium.

Embodiment 3

[0078] The preparation of embodiment 3 in vitro maturation culture medium

[0079] Dissolve FBS, HDACi 4b, CNP, glutamine, penicillin and streptomycin in TCM-199 to prepare a solution containing 15% FBS, 7μM HDACi 4b, 40ng / mL CNP, 0.2mg / mL glutamine, 100IU / mL of penicillin and 100IU / mL of streptomycin in TCM-199 medium is the in vitro maturation medium.

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Abstract

The present invention relates to cell culture, and specifically discloses a culture scheme for immature oocytes, which can promote the in vitro maturation of immature oocytes, specifically comprising follicular granulosa cells and a culture medium for cultivating the follicular granulosa cells, said culture The culture solution of follicular granulosa cells contains CNP or its variant or its analogue and HDAC (histone deacetylase) inhibitor. The present invention further provides an in vitro maturation culture solution comprising CNP or its variants or analogs thereof and HDAC inhibitors and related compositions thereof, as well as the above-mentioned medium, culture solution and composition in promoting the in vitro maturation of immature oocytes Applications.

Description

technical field [0001] The invention relates to an in vitro maturation culture medium for immature oocytes, a related composition and a culture method, and belongs to the field of medicine and biology. Background technique [0002] In human clinical assisted reproduction, gonadotropins are widely used to perform superovulation operations on female patients. However, due to differences in the environment, mental stress, and individual responses to gonadotropins, some patients are undergoing superovulation operations. Finally, it often occurs that a considerable part of the obtained oocytes is immature, which reduces the utilization efficiency of the collected oocytes. Regrettably, the oocyte maturation culture methods and the culture medium used in clinical practice cannot solve the above-mentioned problems to a certain extent. Therefore, if a new method for culturing human immature oocytes in vitro can be developed, so that these immature oocytes can mature and have corresp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/075C12N5/071
CPCC12N5/0609C12N2500/32C12N2501/30C12N2517/10C12N2501/73C12N2502/243C12N2501/065C12N2501/998Y02A50/30C12N5/0018
Inventor 夏国良王超王华荣蔡晗
Owner CHINA AGRI UNIV
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