绒山羊毛囊干细胞的分离培养及诱导分化鉴定

发布时间：2018-08-18 07:36

【摘要】：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12d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。
[Abstract]:The aim of this study was to establish a method for isolation and culture of cashmere goat hair follicle stem cells in vitro and to identify their biological characteristics and multidirectional differentiation potential. Hair follicle stem cells were isolated and purified by two-step enzyme digestion method and differential adherence method. The growth curve and clone formation rate were measured. Oil red O and alizarin red staining were used to identify the adipogenic and osteogenic differentiation of the 5th generation hair follicle stem cells. The results showed that the isolated hair follicle stem cells were homogeneous in morphology, small in volume, typical in paving stone shape, with S shape growth curve and strong clone forming ability. After adipogenic induction, the cell volume increased, the cytoplasm increased, and small lipid droplets appeared in the cell. After 12 days of induction, the lipid droplets increased gradually, and the oil red O staining was positive, and after osteogenesis, the edge of the cells was blurred and granular nodules appeared. It was positive by alizarin red staining. The results showed that the Cashmere Goat hair follicle stem cells had the potential of multiple differentiation.
【作者单位】：内蒙古农业大学生命科学学院;内蒙古自治区生物制造重点实验室;内蒙古农业大学动物科学学院;
【基金】：国家自然科学基金(30901016、31160445)
【分类号】：R329

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