Method for in-vitro separation and culture of goat male germ stem cells

Abstract

The invention discloses a method for in-vitro separation and culture of goat male germ stem cells. Spermatogonia are obtained from the testicular convolutedtubules of a male goat. A method for separating and purifying the goat male germ stem cells adopts 0.2% of gelatin and a Matrigel differential adherence combined cloning method to differentiate and separate un-adhered goat male germ stem cellsfrom other cells for adherence culture; and the culture of the separated goat male germ stem cells is MEF feeder-layer culture of feeder-layer-free culture. The separated goat male germ stem cells have ES cell characteristics and the potential of differentiation into nerve cells, cardiac myocytes and sperm sample 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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