中国组织工程研究与临床康复2009，Vol.13Issue(27)：5286-5290,5.DOI:10.3969/j.issn.1673-8225.2009.27.017
无血清悬浮培养肾癌干细胞表面标志CD133及端粒酶活性的表达
CD133 expression and telomerase activity in serum-free cultured renal carcinoma stem cells
摘要
Abstract
BACKGROUND: Talomerase activity inhibitor inhibits or kills renal carcinoma cells, and also affects stem cells that play importan roles in occurrence and development of renal carcinoma.OBJECTIVE: To observe renal carcinoma stem cell surface marker CD133 and telomerase activity expression in serum-free suspension culture, and to compare with renal carcinoma cells in serum suspension culture.DESIGN, TIME AND SETTING: The in vitro cytological study was performed at the Jiangsu University from June 2008 to Februar 2009.WIATERIALS: Fresh normal renal tissue surrounding renal carcinoma was obtained from Affiliated Hospital, Jiangsu University.Renal carcinoma stem cell line OS-RC-2 was supplied by Cell Bank, Chinese Academy of Sciences Shanghai Branch.METHODS: OS-RC-2 in logarithmic phase, digested by trypsin, and centrifuged. Supematant was removed. OS-RC-2 cell line in serum-free DMEM/F12 supplemented with epidermal growth factor and basic fibroblast growth factor was incubated at 2×105/L in 5% CO2 incubator at 37℃. Renal carcinoma cultured in serum and normal renal tissue served as controls.MAIN OUTCOME MEASURES: Cell growth was observed under an inverted microscope. Expression of CD133 and CD34 was detected using flow cytometry. Reel-time quantitative TRAP assay was applied to evaluate telomerase activity in renal carcinoma stem cells.RESULTS: After incubated in serum-free medium, renal carcinoma stem cells were round and suspended. Two days later, cell mass generated. Each cell mass contained 3-8 cells, with strong refraction. Seven days later, cell mass became more, presented big body that was regular, round or elliptical. CD133+CD34- rate in renal carcinoma stern cell mass was significantly greater in serum-free suspension culture compared with in serum suspension culture. CD133 and CD34 expression was not determined in normal renal tissue. There were significant differences among groups (F=328.25, P < 0.05). Telomerase activity was greater in renal carcinoma stem cells and renal carcinoma cells compared with normal renal ceils (F=-278.74, P < 0.05). No significant difference was detected between renal carcinoma stem cells and renal carcinoma cells.CONCLUSION: Compared with serum cultured renal carcinoma cells, serum-free cultured renal carcinoma cell surface marker CD133 presents high expression. Moreover, talomerase activity is high in renal carcinoma stem cells and renal carcinoma cells compared with normal renal tissue.关键词
肿瘤干细胞/肾癌/端粒酶/活性/CD133分类
医药卫生引用本文复制引用
潘鹏,田福起,郭涛,孙浩,马克钧,周留正..无血清悬浮培养肾癌干细胞表面标志CD133及端粒酶活性的表达[J].中国组织工程研究与临床康复,2009,13(27):5286-5290,5.基金项目
镇江市科技局资助项目(SH2007024)a grant of Science and Technology Bureau of Zhenjiang City, No.SH2007024 （SH2007024）
