健康成人尿源性干细胞的分离、培养与鉴定

Isolation,cultivation and identification of healthy adults urine-derived stem cells

目的:分离培养和鉴定健康成人尿源性干细胞(USCs)。方法:收集3名健康成人新鲜尿液,采用贴壁培养法分离培养细胞。进一步观察细胞形态,对细胞进行染色体核型分析,流式细胞术检测细胞表面标志物CD73、CD90、CD105、CD34和CD45的表达情况,采用茜素红及油红O染色分别检测钙结节和脂质的形成以验证细胞成骨和成脂分化能力。结果:成功从健康成人新鲜尿液中分离到USCs,该细胞贴壁生长,其外观呈长梭形,具有较强的体外增殖能力,且传代过程中其染色体核型并未改变;细胞表面标志物CD73、CD90、CD105的阳性表达率均达到99%,CD34和CD45均呈阴性,阳性率均低于2%;茜素红和油红O染色均呈阳性。结论:从健康成人尿液中成功分离培养出了USCs,并且鉴定了其具有较强的稳定增殖和分化能力。

Objective:To isolate,culture,and identify the urine-derived stem cells(USCs)from healthy adults.Methods:Three fresh urine samples from healthy adults were collected and cells were isolated and cultured by adherent culture.Further,we observed the cell morphology,analyzed the chromosome karyotype,detected the expression of cell surface markers CD73,CD90,CD105,CD34 and CD45 by flow cytometry.Alizarin red and oil red O staining were used to detect the formation of calcium nodules and lipid to verify the ability of osteogenic and adipogenic differentiation.Results:USCs were successfully obtained from the fresh urine of healthy adults.The cells adhered to the plastic,had a long fusiform appearance and a strong ability of proliferation in vitro,and had no change in chromosome karyotype during subculture.The positive expression rates of cell surface markers CD73,CD90 and CD105 were 99%.CD34 and CD45 are negative and positive rates were both less than 2%.Alizarin red and oil red O staining were positive.Conclusion:USCs are successfully isolated and cultured from the urine of healthy adults,and its strong ability of stable proliferation and differentiation is identified.