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Human oocyte in vitro maturation culture solution containing Endothelin-1 and application of human oocyte in vitro maturation culture solution

A technology for in vitro maturation culture and oocyte, which is applied to the in vitro maturation culture medium of human oocytes and the application field of in vitro maturation culture of human immature oocytes, which can solve problems such as ET-1 that have not yet been seen.

Active Publication Date: 2015-11-04
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] So far, there has been no report on the application of ET-1 in the field of in vitro maturation of human oocytes

Method used

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  • Human oocyte in vitro maturation culture solution containing Endothelin-1 and application of human oocyte in vitro maturation culture solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment one : Preparation of human oocyte maturation medium containing ET-1 in vitro

[0024] Liquid A: weigh ET-1 10 mu g, 7.5IU r-FSH, 10ml SSS, dilute to 100ml with G-IVF, mix well, use 0.22 mu m microporous filter membrane (Millipore) and stored in a refrigerator at 4°C.

[0025] Liquid B: weigh ET-1 10 mu g, 7.5IU r-FSH, 10ml SSS and 50IU hCG, dilute to 100ml with G-IVF, mix well, use 0.22 mu m microporous filter membrane (Millipore) and stored in a refrigerator at 4°C.

Embodiment 2

[0026] Embodiment two : Human oocyte in vitro maturation medium containing ET-1 is used for in vitro maturation of human oocytes

[0027] Human immature oocyte-cumulus complexes (OCC) were divided into three groups, respectively in G-IVF culture medium (control group), containing 0.075IU / ml r-FSH, 0.5IU / ml hCG, volume fraction 10 %SSS G-IVF culture medium and the human oocyte in vitro maturation medium containing ET-1 prepared in Example 1 were used for in vitro maturation culture, followed by ICSI in vitro fertilization. The specific method is as follows:

[0028] 1. Reagent balance preheating: Add G-IVF culture solution (control group) containing 10% SSS by volume fraction, 0.075IU / ml r-FSH, 0.5IU / ml HCG to different four-well petri dishes, volume G-IVF culture medium with a fraction of 10% SSS and the culture medium for in vitro maturation of human oocytes containing ET-1 prepared in Example 1. In particular, it should be pointed out that the addition method of human oo...

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Abstract

The invention provides a human oocyte in vitro maturation culture solution containing Endothelin-1. The human oocyte in vitro maturation culture solution comprises liquid A and liquid B, wherein the liquid A comprises ET-1, r-FSH and 10% SSS, the liquid B comprises ET-1, r-FSH, hCG and 10% SSS, and solvent is conventional basic culture solutions. Researches show that the culture solution and culture method are capable of promoting human oocyte in vitro maturation especially cytoplasmic maturation and can increase embryo development potential after human oocyte in vitro fertilization. The culture solution aiming at the current situation that development imbalance of cytoplast and cell nucleus during immature human oocyte in vitro culture affects embryo in vitro development potential is developed by utilizing the biological function and effect of ET-1 in cumulus cells. The culture solution has the advantages that the culture solution is applicable to human oocyte in vitro culture, an IVM process can be optimized favorably, and the culture solution is worthy of popularization and application.

Description

technical field [0001] The invention belongs to the biomedical technical field of reproductive medicine, and relates to an in vitro maturation culture medium for human oocytes containing Endothelin-1 (ET-1) and its application in in vitro maturation and cultivation of immature human oocytes. Background technique [0002] Human oocyte maturation in vitro (in vitro maturation, IVM) refers to the development of immature oocytes in follicles into mature second meiotic metaphase (MII) oocytes through in vitro culture, fertilization and division into embryos And transplant the technology to obtain pregnancy. Compared with conventional in vitro fertilization-embryo transfer (in vitro fertilization-embryo transfer), IVM can reduce the cost of high-dose ovulation-stimulating drugs, save time for medical treatment, and more importantly, avoid side effects caused by drugs to stimulate ovaries. In some fields of reproductive medicine, IVM has an irreplaceable role, such as: IVM for pat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075
Inventor 叶英辉崔龙曲凡
Owner ZHEJIANG UNIV
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