人尿源干细胞的提取和鉴定

Isolation and characterization of human urine stem cells

目的从正常人尿液中分离得到具有增殖和分化能力的人尿源干细胞(h USC)。方法收集16例健康成人新鲜尿液,分离培养得到贴壁细胞,显微镜观察拍照。WST-1检测并绘制细胞生长曲线。流式细胞术分析细胞表面分子表达率。成骨和成脂诱导培养基诱导h USC分化。结果采用优化的细胞培养基得到12株具有较快增殖能力的h USC。分离的细胞中表达CD13、CD29、CD73、CD105的细胞数目均大于97﹪,表达CD90的细胞数目不足50﹪,表达造血系表面抗原CD14、CD19、CD34、CD45,内皮细胞表面抗原CD31以及HLA-DR的细胞数目均小于1﹪。h USC经诱导可分化成为骨和脂肪细胞。结论采用本研究自建培养体系可培养得到形态单一、具有较强增殖分化能力的人尿源干细胞。

Objective To isolate cells from human urine samples and determine whether the cells could be expanded and differentiate into mesoderm cells. Method Fresh urine samples were coilected from 15 healthy individuals. Urine cells were isolated and expanded. Cell morphology was observed using a microscope. The proliferation of cultured urine cells was determined by water-soluble tetrazolium salt （WST-1） assay. Cell surface markers were detected using flow cytometry. Osteogenic and adipogenic medium was used to induce human urine stem cells into osteogenic and adipogenic lineages. Result Cells were obtained from 12 urine samples using our USC cell medium. FACS revealed that urine cells were positive for cell surface markers CD13, CD29, CD73, and CD105. Almost half of the hUSCs expressed CD90. The hUSCs were negative for the hematopoietic cell markers CD14, CD19, CD34, and CD45, endothelial lineage markers CD31, and HLA-DR. Urine cells also had the potential to differentiate into osteogenic and adipogenic cell lineages. Conclusion In this study, our hUSC cell medium can be used to isolate urine cells, which have proliferation and differentiation abilities.