A kind of in vitro isolation and culture method of goat male reproductive stem cells

Abstract

The invention discloses an in vitro separation and culture method of goat male reproductive stem cells. The spermatogonia are obtained from the seminiferous tubules of the testes of male goats. The separation and purification method of goat male reproductive stem cells adopts 0.2% gelatin and Matrigel differential adhesion Combined with the cloning method, the non-adhered goat male reproductive stem cells were distinguished and separated from other cells adhered to the wall. The culture of the isolated goat male reproductive stem cells is MEF culture with feeder layer or culture without feeder layer. The isolated goat male reproductive stem cells have the characteristics of ES cells and the differentiation potential to nerve cells, cardiomyocytes 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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