SymbolCD133 expression and telomerase activity in serum-free cultured renal carcinoma stem cells
- Pan, Peng
- Tian, Fu-qi
- Guo, Tao
- Sun, Hao
- Ma, Ke-jun
- Zhou, Liu-zheng
Department of Urology Surgery, Affiliated Hospital, Jiangsu University, Zhenjiang 212001, Jiangsu Province, China
Pan Peng, Associate chief physician, Department of Urology Surgery, Affiliated Hospital, Jiangsu University, Zhenjiang 212001, Jiangsu Province, China [email protected]
Correspondence to: Guo Tao, Department of Urology Surgery, Affiliated Hospital, Jiangsu University, Zhenjiang 212001, Jiangsu Province, China [email protected]
Supported by: a grant of Science and Technology Bureau of Zhenjiang City, No. SH2007024*
Received: 2009-04-07
Accepted: 2009-05-21
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Abstract
BACKGROUND:
Telomerase activity inhibitor inhibits or kills renal carcinoma cells, and also affects stem cells that play important 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 February 2009.
MATERIALS:
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. Supernatant 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 2×108/L in 5% CO2 incubator at 37 °C. 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. Real-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 stem 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 cells (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, telomerase activity is high in renal carcinoma stem cells and renal carcinoma cells compared with normal renal tissue.
Pan P, Tian FQ, Guo T, Sun H, Ma KJ, Zhou LZ. CD133 expression and telomerase activity in serum-free cultured renal carcinoma stem cells.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2009;13(27): 5286-5290. [http://www.crter.cnhttp://en.zglckf.com]
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