In-vitro separation and preparation method of male germline stem cells of goat

Abstract

The invention discloses an in-vitro separation and preparation method of male germline stem cells of a goat. Spermatogonium can be obtained from male convoluted tubules of the goat. The separation and preparation method of germline stem cells of a male goat comprises the following steps: carrying out differential adhesion by using 0.2% gelatin and Matrigel through combining a cloning method; and distinguishing and separating non-adhesion male germline stem cells of the goat with other adhesion culture cells, wherein the culture of separated male germline stem cells of the goat is MEF (mouse embryonic fibroblast) feeder culture or feeder-free culture. The separated male germline stem cells of the goat have embryonic stem (ES) cell features and proficiency of differentiating to be nerve cells, cardiocytes and sperm-like cells.

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 goat male 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 sperm cell generation and maturation disorders is a real problem that plagues many patients; In research, sperm is one of the most important research objects of developmental biology research; therefore, the research on the occurrence and differentiation of germ cells is of great significance in animal husbandry production and biomedical research. However, the occurrenc...

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Problems solved by technology

However, people still don't know enough about the reproductive stem cells of large livestock such as cattle, sheep, and pigs. The only few research reports that have been seen involve the isolation and culture of mGSCs cells, but there is no standardized isolation and culture method for reproductive stem cells (Dong Wuzi et al., In vitro isolation and culture of goat fetal testicular cells and the growth behavior of male germline stem cells. Journal of Agricultural Biotechnology, 2004, 12(6): 648-652.; Dong Wuzi et al., Isolation of ES cells derived from fetal bovine male reproductive stem cells Cultivation and pluripotency research. Chinese Journal of Bioengineering, 2007, 23(4): 751-755; Dong Wuzi et al., Culture and detection of ES-like cells derived from newborn bovine male germ cells. Journal of Animal Husbandry and Veterinary Medicine, 2007, 38(2): 144-148)

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