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In-vitro separating and culturing method for germline stem cell

A technology for the separation and cultivation of reproductive stem cells, which is applied in the direction of artificial cell constructs, animal cells, vertebrate cells, etc., can solve the problems that are insufficient and have not yet been seen in the separation and cultivation of female reproductive stem cells, and achieve large nuclei, translucent cells, and light-shielding strong effect

Active Publication Date: 2011-11-23
陕西九州细胞基因工程有限公司
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Problems solved by technology

However, the reproductive stem cells of domestic animals such as pigs, cattle, and sheep are still not well understood, and there is no method for isolating and culturing female reproductive stem cells.

Method used

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  • In-vitro separating and culturing method for germline stem cell
  • In-vitro separating and culturing method for germline stem cell
  • In-vitro separating and culturing method for germline stem cell

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Embodiment Construction

[0034] The invention provides a culture system for efficiently isolating and cloning mammalian female reproductive stem cells, including the separation of female reproductive stem cells, detection and in vitro culture of female reproductive stem cells.

[0035] The adult mammalian ovary is used as the source of raw materials; the isolation and purification method of female reproductive stem cells adopts 0.2% gelatin and Matrigel differential attachment binding cloning method to distinguish and separate non-adherent female reproductive stem cells from other cells adhered to the wall.

[0036] The in vitro separation of pig female reproductive stem cells is specifically described below:

[0037] A, the in vitro separation method of pig female reproductive stem cells, specifically comprising the following steps:

[0038] 1) Isolation of porcine ovary cells

[0039] The aseptically collected porcine ovaries were washed 3 times with sterile saline containing double antibodies (500...

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Abstract

The invention discloses an in-vitro separating and culturing method for a germline stem cell. The method comprises the following steps of: obtaining a cell from germinal epithelium of an ovary of a female mammal, differentiating and separating a non-adherent germline stem cell from other adherent cultured cells by adopting gelatin with the concentration 0.2 percent and a Matrigel differential adherence combination cloning method; and culturing the germline stem cell obtained by separating on an MEF (Mouse Embryonic Fibroblast) feeder layer. The separated germline stem cell has ES (Embryonic Stem) characteristic and the potential of differentiating into a muscle cell, an adipose cell and oocyte-like cell and various cells with three internal germinal layers.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the in vitro proliferation, differentiation and separation of animal stem cells, in particular to a method for in vitro separation and cultivation of female reproductive stem cells. Background technique [0002] The research on the occurrence, proliferation and differentiation of germ cells is one of the important topics in life science research. In animal breeding and breeding, high-yield and high-quality genetic resources are completely dependent on germ cells to complete heredity; in humans, infertility caused by ovum generation and maturation disorders is a real problem that plagues many patients; in basic research Among them, ovum is one of the most important research objects of developmental biology research; therefore, the research on the occurrence and differentiation of oocyte is of great significance in animal husbandry production and biomedical research. However, the occurr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 华进联胡玥白耀富
Owner 陕西九州细胞基因工程有限公司
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