【摘要】 目的体外分离、纯化及培养人脐带间充质干细胞(UC-MSCs),观察其生物学特性并研究其定向分化成汗腺样细胞的能力。方法将剔除动静脉的新鲜人脐带组织切成小段,剥离血管后酶解,释放细胞贴壁生长,倒置显微镜观察细胞形态,绘制第3代生长曲线;流式细胞仪测定细胞表面标记(CD105、CD73、CD90、CD34、CD45、CD14和HLA-DR);定向诱导分化后PT-PCR检测汗腺发育基因EDA及EDAR表达。结果原代人脐带间充质干细胞呈现典型的成纤维样形态。体外诱导实验证实,该细胞阳性表达汗腺特异基因。结论人脐带MSCs体外可以分化成汗腺样细胞,为利用人脐带间充质干细胞修复汗腺损伤提供了可靠地理论依据和实验基础。更多还原

【Abstract】 Objective To isolate, purify and culture human umbilical cord mesenchymal stem cells(UC-MSCs) in vitro, and to observe the biological characteristics of human umbilical cord mesenchymal stem cells(MSCs) and to study their ability to differentiate into sweat gland like cells. Methods Cut into small pieces of arteriovenous excluding fresh human umbilical cord tissue, artery dissection after enzymatic hydrolysis, releasing adherent cell growth, inverted microscope observation cell morphology, draw a third generation growth curve, flow cytometry determination of cell surface markers(CD105, CD73, CD90, CD45, CD34, CD14 and HLADR), directional differentiation of RT-PCR sweat glands development gene of EDA and EDAR expression. Results After inoculation of the third generation of cells, which <72 h incubation period, logarithmic growth phase was 72 ～96 h, when to 7 ～ 8 d was for growth plateau. In this study, the results showed that there was no obvious cell aging and the proliferation rate of the 15 generations. Conclusion Human umbilical cord MSCs can differentiate into sweat gland like cells in vitro, which provides a reliable theoretical basis and experimental basis for the use of human umbilical cord mesenchymal stem cells in the repair of sweat gland injury.更多还原