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In vitro maturation medium and application of human oocytes containing endothelin-1

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

Active Publication Date: 2018-08-28
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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  • In vitro maturation medium and application of human oocytes containing endothelin-1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0024] Solution A: Weigh 10μg of ET-1, 7.5IU r-FSH, 10ml of SSS, dilute to 100ml with G-IVF, mix well, use 0.22μm microporous filter membrane (Millipore) to filter and store in 4℃ refrigerator .

[0025] Solution B: Weigh 10μg of ET-1, 7.5IU r-FSH, 10ml SSS and 50IU hCG, use G-IVF to make up to 100ml, mix well, use 0.22μm microporous filter membrane (Millipore) to filter and store in 4°C refrigerator.

Embodiment 2

[0026] Embodiment 2: Human oocyte in vitro maturation medium containing ET-1 is used for in vitro maturation of human oocyte

[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 noted that the human oocyte maturation medium containi...

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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-embryotransfer), IVM can reduce the cost of high-dose ovulation-stimulating drugs, save time for medical treatment, and more importantly, avoid the side effects of drugs to stimulate ovaries. In some fields of reproductive medicine, IVM has an irreplaceable role, such as: IVM for patient...

Claims

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Application Information

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