绒山羊毛囊干细胞的分离培养及诱导分化鉴定

试验旨在建立绒山羊毛囊干细胞体外分离培养方法,并对其生物学特性和多向分化潜能进行鉴定。利用两步酶消化法和差速贴壁法分离纯化毛囊干细胞,对其进行传代培养,测定生长曲线和克隆形成率,采用油红O和茜素红染色法对第5代毛囊干细胞向成脂和成骨分化情况进行鉴定。结果显示,分离得到的毛囊干细胞形态均一,体积小,呈典型的铺路石状生长,生长曲线呈S型,克隆形成能力强。成脂诱导后,细胞体积增大,细胞质增多,胞内出现小脂滴,诱导12 d后,脂滴逐渐增多,油红O染色呈阳性;成骨诱导后,细胞边缘模糊,出现颗粒状结节,经茜素红染色呈阳性。表明获得的绒山羊毛囊干细胞具有多向分化的潜能。

Isolation and Cultures of Hair Follicle Stem Cell from Cashmere Goat and Identification of Induced Differentiation

The aims of this paper were to explore the optimal method of isolating,purifying,and proliferating the Cashmere goat hair follicle stem cells,and their multiple differentiation potentiality. It separated and cultivated the Cashmere goat hair follicle stem cells by two methods,one was the two-step enzyme digestion,another was differential adhesion. Growth curves and colony forming efficiency of passaged the Cashmere goat hair follicle stem cells were determined. The P5 hair follicle stem cells were cultured in vitro under adipogenic and osteogenic inductive environment and induced into adipocytes,osteoblates. The results showed that the isolated hair follicle stem cells were uniform,volume small,paving stone-like,and active proliferative capacity and had S shape of growth curve. Their adipogenic and osteogenic differentiation properties were confirmed by histological staining such as oil red O and alizarim red. Oil red O and alizarim red staining of adipogenic and osteogenic induction group was positive,whereas,control groups were negative.