体外人骨髓间充质干细胞向汗腺细胞表型转化的实验研究

An experimental study of conversion of human bone marrow mesenchymal stem cells into phenotype of sweat gland cells in vitro

目的:研究骨髓间充质干细胞(MSCs)向汗腺细胞分化的可行性。方法:体外分别分离培养、扩增并鉴定MSCs和汗腺细胞,将汗腺细胞置于47℃环境中1 h建立汗腺细胞体外休克模型,继续孵育3～5 d,收集上清液作为条件培养基对MSCs分化诱导,应用免疫组织化学和流式细胞仪法检测对比共培养10 d后MSCs细胞表型的改变。结果:检测MSCs表面标志CS29、CD44、CD105阳性,汗腺细胞表面标志角蛋白(CK)7、CK8、CK18、CK19、CEA阳性;MSCs诱导分化后CEA、CK7、CK8和CK19染色阳性。而未诱导的MSCs没有检测到CEA、CK7、CK8和CK19染色阳性的细胞。结论:通过建立适当的体外汗腺诱导培养体系,中胚层的MSCs可以通过跨胚层分化途径转变为汗腺样细胞。

Objective： To investigate the feasibility of differentiation of human bone marrow mesenchymal stem cells （MSCs） to sweat gland like cells. Methods： MSCs and sweat gland cells were isolated, cultured, expanded and identified in vitro respectively. The sweat gland cells were cultured at 47 ℃ for 1 hour. The sweat gland cells cultured in the fresh sweat gland cell culture medium served as controls. After continuous culturing for 3-5 days at normal temperature, supernatant was collected as conditioned medium for inducing differentiation of MSCs. Using immunohistochemistry and flow cytometry techniques to determine morphological changes of MSCs after co-culture for 10 days. Results： CD29, CD44, CD105 were positive in MSCs, and CK, CK8, CK18, CD19, CEA were positive in sweat gland cells. Cells positive for CEA, CK7, CK8 and CK19 were found when MSCs were induced in conditioned medium. MSCs were negative for CEA, CK7, CK8 and CK19 in control group. Conclusions： MSCs are induced successfully and cells with the phenotype of sweat gland cells were found. By the cross-mesoderm pathway, mesodermal MSCs can be converted into sweat gland-like cells by co-culture with suitable culture system.