成年山羊皮肤干细胞体外克隆与诱导分化

CLONING AND INDUCED DIFFERENTIATION OF ADULT GOAT SKIN STEM CELLS IN VITRO

目的 探索山羊皮肤干细胞的分离方法与培养体系 ,为进一步研究皮肤干细胞增殖分化的分子机制打下基础。 方法 从成年关中奶山羊耳尖获取皮肤 ,用组织块培养法分离皮肤干细胞 ,碱性磷酸酶 (AL P)染色及体外分化实验鉴定 ,用条件培养基与细胞因子相结合行皮肤干细胞的体外诱导分化。 结果 山羊耳尖皮肤分离得到类干细胞集落并传 5代 ,细胞集落具有典型鸟巢状结构 ,AL P染色阳性 ,体外分化能形成类胚体。在细胞因子 (IGF- 1)及条件培养基的诱导下 ,所分离获取的皮肤干细胞能分化形成类毛囊 ,类星形胶质细胞及类成骨细胞。 结论 用组织块培养法可以获得多能性皮肤干细胞 ;在体外培养条件下不仅能进行定向分化 ,还能进行横向分化。

Objective To establish a good method and culture system to isolate skin stem cell and expand it in vitro so as to lay a foundation for exploring the proliferation and differentiation mechanism of skin stem cell. Methods Skin stem cells were obtained by explant culture and identified by using alkaline phosphatase(ALP) staining and differentiating experiment in vitro. Stem cell was induced by the cocktail of conditional medium with cell growth factor (insulin like growth factor and epidermal growth factor). Results Skin stem cell colonies were derived from ear skin tissues of adult dairy goats. The colonies had some characteristics of embryonic stem cells, such as the ability to be continuously passaged (Passage 5) and the morphology nest like. They continued to be ALP positive and had the capacity of forming embryoid bodies. These cells were pluripotent and stem like cells. In vitro these stem cell can be induced to be Follicle like structure, Astrocyte like cells, osteoblast like cell. Conclusion Explant culture is a good method to isolate skin stem cell, which can be induced to be committed differentiation and trans differentiation.