Abstract: The experiment was a preliminary study on fine-wool sheep hair follicle stem cells, which were isolated and cultured, in order to establish the fine-wool sheep hair follicle stem cells culturing system in vitro. It separated and cultivated the fine wool sheep hair follicle stem cells by three methods, one was the two-step enzyme digestion, another one was mechanical separation, and the third was the combination of two-step enzyme digestion and mechanical separation. By measuring the number, cloning efficiency and proliferation ability of hair follicle stem cells, the experiment intended to evaluate the advantages and disadvantages of the three methods synthetically, for not only obtaining a large number of hair follicle stem cells conveniently, but also seeking out ideal culture conditions of reducing their differentiation. Under the inverted microscope, the cells obtained by three cultural methods were paving stone-like, by which expressed keratin K19 and integrin-β1, indicating successful acquirement of sheep hair follicle stem cells and establishment of in vitro culture system. Finally, the experiment showed that, it was the optimal method to obtain and culture hair follicle stem cells by the combination of two methods of two-step enzymatic digestion and mechanical separation.

Key words: hair follicle stem cells; culture in vitro; proliferation capacity