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Method for promoting primordial follicle library formation by activin A

A primordial follicle and activator technology, applied in the fields of biochemical equipment and methods, microbial determination/testing, germ cells, etc.

Inactive Publication Date: 2014-03-26
QINGDAO AGRI UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

At present, there is no research report on the role of activin A in regulating the formation of primordial follicle pool of germ cells before meiosis

Method used

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  • Method for promoting primordial follicle library formation by activin A
  • Method for promoting primordial follicle library formation by activin A
  • Method for promoting primordial follicle library formation by activin A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The operation steps of activin A of the present invention to promote the formation of primitive follicle bank are as follows:

[0020] 1) Using in vivo and in vitro experiments to determine that the critical period for activin A to play a role is from 12 days before birth to 7 days after birth: the fetal mouse ovarian tissue of 12.5dpc was mechanically isolated and kept at 37°C and 5% CO 2, cultured in an incubator with saturated humidity, half of the culture medium was changed every two days, culture medium A contained 88% (α-MEM), 10% fetal calf serum (FCS), 1% sodium pyruvate, 1% streptomyces lividans 10mIU follicle-stimulating hormone (FSH); the culture was divided into four stages, 7 days in each stage, a total of 28 days; in each stage, a group without activin A was set up, and when the culture reached 28 days, the fetal mouse ovaries were collected and torn. Broken into small tissue pieces, treated with 0.25% trypsin and 0.2% collagenase IV at 37°C for 10 min, ad...

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Abstract

The invention relates to a method for promoting primordial follicle library formation by activin A, and the steps are as follows: 1) using in-vivo and in-vitro experiments to determine that the critical period for the activin A to play a role is 12 days before birth to 7 days after the birth; 2) using cell fluorescence staining and western blot to detect the expression of meiosis related proteins to determine activin-A-promoted first meiotic process of female reproductive cells; 3) using a fluorescent immunohistochemistry method for statistics of primordial follicle formation situation in each group, wherein the activin A promoting the primordial follicle library formation. The method has the advantages of simple operation and reliable principle, the activin A plays an important role in early oocyte occurring stage, the activin A enables the retinoic acid concentration in the female genital ridge to remain relatively high, and retinoic acid promotes the expression of Stra8 gene in the female reproductive cells, accelerates meiotic starting and process of the female reproductive cells, and promotes the primordial follicle formation.

Description

Technical field: [0001] The present invention relates to a method for activin A (Activin A) to promote the first meiotic process of female germ cells, thereby promoting the establishment of primordial follicle bank in mammals, in particular to a method for activin A to promote the formation of primordial follicle bank method. It belongs to the field of biomedical technology of reproductive medicine. Background technique: [0002] With the rapid development of global industrialization and the continuous aggravation of environmental pollution, the proportion of infertile couples is gradually increasing, and the incidence of fetal birth defects remains high. The incidence of diseases such as female infertility, premature ovarian failure and congenital birth defects in China is also gradually increasing. Researchers have conducted a lot of research on the causes of infertility and birth defects and found that one of the main causes of these ovarian reproductive diseases is the...

Claims

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Application Information

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IPC IPC(8): C12N5/075C12Q1/68G01N33/68G01N33/53
Inventor 梁桂金刘京才赖方秾秦训思程顺峰沈伟
Owner QINGDAO AGRI UNIV
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