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Immature oocyte in-vitro maturation culture solution and application thereof

A technology for in vitro maturation and culture of oocytes, applied in cell culture media, cell culture active agents, artificial cell constructs, etc., can solve problems such as unsatisfactory in vitro culture systems for oocytes, improve in vitro development ability, increase The probability of successful conception, the effect of promoting expression and secretion

Active Publication Date: 2017-12-15
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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

Method used

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  • Immature oocyte in-vitro maturation culture solution and application thereof
  • Immature oocyte in-vitro maturation culture solution and application thereof
  • Immature oocyte in-vitro maturation culture solution and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] The preparation of embodiment 1 in vitro maturation culture medium

[0084] 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

[0085] The preparation of embodiment 2 in vitro maturation culture medium

[0086] 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

[0087] The preparation of embodiment 3 in vitro maturation culture medium

[0088] 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 invention relates to cell culture and particularly discloses an immature oocyte culture scheme which can promoting in-vitro maturation of immature oocytes. Specially, the scheme adopts follicular granulosa cells and a culture solution for culturing the follicular granulosa cells, wherein the culture solution for culturing the follicular granulosa cells contains CNP or its variants or analogues and an HDAC (histone deacetylase) inhibitor. The invention further provides an in-vitro maturation culture solution containing the CNP or its variants or analogues and the HDAC inhibitor and related compositions and application of a culture medium, the culture solution and the compositions in promotion of in-vitro maturation of immature oocytes.

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, and then use the obtained mature oocytes to perform in vitro fertilization or single sperm injection to achieve the goal of conceiving offspring. However, due to environmental, mental stress, and individual differences in response to gonadotropins, some patients often experience a considerable portion of immature oocytes after performing superovulation operations, which greatly reduces the risk of ovulation. The success rate of assisted reproduction in this patient decreased the utilization efficiency of the collected oocytes. Regrettably, the oocyte maturation culture methods and the culture medium used in clinical prac...

Claims

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Application Information

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Patent Type & Authority Applications(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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