毛囊干细胞细胞膜片的构建研究

Study on Construction of Cell Membrane of Hair Follicle stem Cells and Research on Its Structure

目的以毛囊干细胞作为种子细胞,构建细胞膜片,检测培养的细胞膜片的结构,为进一步进行动物试验奠定基础。方法采用中性蛋白酶(Dispase)分离人毛囊干细胞,构建细胞膜片。该膜片构建成功后,分别进行扫描电镜观察、HE染色、角蛋白15和角蛋白10免疫组织化学染色。结果经过1～2周的培养,细胞连接成片,呈现典型的"铺路石"状。将细胞膜片消化下来,HE染色显示该膜片由2～4层细胞组成,电镜观察示细胞排列紧密,细胞基质分泌多。免疫组织化学染色显示细胞膜片大部分细胞K15染色阳性,少量细胞K10染色阳性。结论由毛囊干细胞构建的细胞膜片与人正常表皮结构类似,可进一步深入研究。

Objective To use the hair follicle stem cells as the seeding cells to construct the cell membrane, then to examine its structure for the purpose of the further graft experiment. Methods The dispase was used to isolate the cell membrane. After the cell membrane was constructed, it was examined by transmission electron microscope, HE staining and immunhistochemical staining with monoclonal antibody against cytokeratin 15 （K15） and cytokeratin 10 （K10）. Results The cells could grow confluent after 1 to 2 weeks and were like the slab stone. After the cell membrane was digested, HE staining shows the cell membrane was made up of 2 to 4 layers of cells. Transmission electron microscope examination found the cells arranged closely, the matrix secreted abundantly. Immunohistochemical examination demonstrated that the most cells of the membrane expressed keratinlS, the small fraction of the cells expressed keratin10. Conclusion The cell membrane constructed by hair follicle stem cells is similar to the normal epidermis in the structure, and it can be used as the epidermis substitute in the further study.