人类尿液来源干细胞的体外培养和生物学特性分析  

In vitro Culture and Biological Characteristics of Urine-Derived Stem Cells

作　　者：牛鑫[1] 张国威[2] 汪泱[1] 

机构地区：[1]上海交通大学附属第六人民医院四肢显微外科研究所,上海200233 [2]上海交通大学附属第六人民医院神经外科,上海200233

出　　处：《实验与检验医学》2017年第5期638-641,共4页Experimental and Laboratory Medicine

基　　金：国家自然科学基金(No.81501863);上海市第六人民医院院级课题(No.1634)

摘　　要：目的从人类尿液中直接分离能够体外大量扩增且具有多向分化潜能的干细胞,并研究其生物学特性。方法收集志愿者尿液100~200ml,经离心后使用完全培养基重悬,接种于培养板中;克隆贴壁生长后换液。大量扩增后通过流式细胞术检测其表面标志物,使用成骨、成软骨诱导培养基对其进行诱导分化,通过茜素红染色、阿利新蓝染色检测诱导分化结果。结果每100ml尿液经14d培养能够直接分离得到3~10个克隆,传代培养至第5代时可扩增至2~10×10~7个尿液来源干细胞,能够满足细胞治疗需要。尿液来源干细胞表面标志物CD29,CD44,CD73,CD90为阳性,CD34,CD45,HLA-DR为阴性,与间充质干细胞一致。经诱导培养后,尿液干细胞能够能够成骨、成软骨分化。结论尿液来源干细胞具有间充质干细胞特性,可以作为细胞治疗、组织工程中的种子细胞。Objective To directly isolate in vitro proliferative and multipotential stem cells from urine specimen and study their biological characteristics. Methods 100-200ml urine were collected from volunteers,and further to centrifuged urine and resuspended the precipitate,then seeded the suspension into tissue culture plates. Changed fresh media after clones appeared. Detected cell surface markers through flow cytometry. Osteogenic and chondrogenic induction were performed followed by Alizarin Red S staining and Alcian Blue staining. Results We could isolated 3-10 clones from 100 ml urine after cultured for 14 days. After subculturing to the fifth generation we harvest 2 -10 ×10^7cells. These urine-derived stem cells expressed CD29,CD44,CD73 and CD90,but not expressed CD34,CD45 and HLA-DR,which is consistent with mesenchymal stem cells. Conclusion Urine-derived stem cells have similar biological characteristics with mesenchymal stem cells,and could act as seed cells for cell therapy and tissue engineer.

关 键 词：尿液来源干细胞 间充质干细胞 种子细胞 

分 类 号：Q813.11[生物学—生物工程] R446.62[医药卫生—诊断学]