尿源性干细胞的提取及鉴定

Extraction and identification of human urine stem cells

目的:对人尿源性干细胞(hUSCs)进行分离提取和鉴定,为干细胞的应用打下基础。方法:无菌操作下收集2例年龄20~40岁的健康成人约200 mL新鲜中段尿,离心法初步提取尿液细胞,用干细胞培养液筛选培养hUSC,细胞传代至第3代时冻存,流式细胞技术鉴定第4代hUSCs表面标志物,用成脂试验检测干细胞的诱导分化能力。结果:2例均提取成功,在培养第7天时成功提取出hUSCs,并体外培养形成集落,流式细胞术鉴定干细胞表面抗原CD44、CD105和CD73呈高表达,CD45和HLA-DR呈低表达,表明具有间充质干细胞的特性,成脂试验检测干细胞具有诱导分化能力。结论:运用特殊的干细胞培养基成功培养出hUSCs并保存,可建立成熟稳定的培养技术和体系。

Objective:To isolate,extract,and identify human urinary derived stem cells(hUSCs),laying a foundation for the application of stem cells.Methods:Two healthy adults aged between 20 and 40 years old,approximately 200 mL of fresh midstream urine,were collected under aseptic operation.Urine cells were initially extracted by centrifugation,and hUSCs were screened and cultured using stem cell culture medium.Cells were passaged to the third generation and frozen.Surface markers of the fourth generation human urine derived stem cells were identified by flow cytometry,and their ability to induce differentiation was tested using adipogenesis assay.Results:Both cases were successfully extracted,and hUSCs were successfully extracted on the 7th day of cultivation and cultured in vitro to form colonies.Flow cytometry identified high expression of stem cell surface antigens CD44,CD105and CD73,while low expression of CD45 and HLA-DR indicated the characteristics of mesenchymal stem cells.The specific induction of differentiation ability of stem cells was detected by adipogenesis assay.Conclusion:hUSCs were successfully cultured and preserved using special stem cell culture media,and a mature and stable culture technology and system were established.