摘 要 ： 为建立高效简便的人尿源干细胞（Human urine-derived stem cells，hUSCs）培养和鉴定体系，在无菌条件下收集青年志愿者的尿液分离培养人尿源干细胞，然后用克隆计数、形态观察、核型分析、生长曲线测定、表面标志物检测、成骨和成脂分化等方法对 hUSCs 进行鉴定。人尿源干细胞克隆具有纤维样、鹅卵石样和丝连样三种形态。第二代 hUSCs 高表达抗原CD44、CD73和CD90，阳性率均高于99.6% ；而低表达抗原CD34 和 CD45，阳性率均低于0.3%。第六代hUSCs高表达抗原 CD44和CD73，阳性率均高于 99.5% ；而低表达抗原 CD34 和 CD45，阳性率均低于 0.5%。第六代 hUSCs 仍然具有很强的增殖能力和稳定的核型，并经诱导可分化为骨细胞和脂肪细胞。该研究建立了高效简便的 hUSCs 培养和鉴定体系。

　　关键词 ：人尿源干细胞；生长曲线 ；表面标志 ；种子细胞 ；再生医学

　　DOI ：10.13560/j.cnki.biotech.bull.1985.2018-0312

　　WANG Fu-ping1，2 ZHANG Feng1，ZHAO Tong-biao2

　　（1.School of Life Science，Hebei University，Baoding 071002 ；2.Institute of Zoology，Chinese Academy of Science，Beijing 100101）

　　Abstract: In order to establish an efficient and convenient system for the cultivation and identification of human urine-derived stem cells（hUSCs），urine samples from young volunteers were collected under aseptic conditions to isolate and culture hUSCs. The clone assay，morphological observation，karyotype analysis，growth curve determination，surface marker detection，osteogenic and adipogenic differentiation were utilized to identify the hUSCs. The clones of hUSCs were in three types of fiber-like，pebble-like and silk-like. For the second passage hUSCs highly-expressed antigens CD44，CD73 and CD90，the positive rates were higher than 99.6% ；while for the lowlyexpressed antigens CD34 and CD45，the positive rates were less than 0.3%. For the sixth passage hUSCs highly-expressed antigens CD44 and CD73，and the positive rates higher than 99.5%；while for the lowly-expressed antigens CD34 and CD45，the positive rates were less than 0.5%. The sixth passage hUSCs still had strong proliferative ability and stable karyotype，and was induced to differentiate into osteocyte and adipocyte. By this research，the efficient and convenient hUSCs culture and identification systems are established.

　　Key words: human urine-derived stem cells ；growth curve ；surface marker ；seed cell ；regenerative medicine