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目的:研究分析人肾癌干细胞的培养鉴定及其端粒酶活性情况。方法:应用含有碱性成纤维生长因子(英简b FGF)、表皮生长因子(英简EGF)的DMEM/F12的培养基对分离的肾癌干细胞进行无血清悬浮培养,并以正常的肾组织细胞及含血清培养的肾癌细胞作为对照。通过端粒重复序列扩增法(英简TRAP)对肾癌干细胞端粒酶活性进行实时、定量的检测。结果:无血清悬浮培养组的CD133~+CD34~+的表达率显著高于含血清培养组,差异P<0.05有统计学意义。并且,肾癌干细胞球、肾癌细胞端粒酶活性均显著高于正常肾组织细胞,差异P<0.05有统计学意义。结论:无血清悬浮培养相比含血清培养,肾癌干细胞CD133~+的表达率更高,但二者培养的肾癌细胞的端粒酶活性均明显高于正常肾组织细胞。
Objective: To study the culture identification and telomerase activity of human renal cancer stem cells. Methods: The isolated renal carcinoma stem cells were cultured in serum-free medium containing DMEM / F12 containing basic fibroblast growth factor (FGF) and epidermal growth factor (EGF), and cultured in normal renal tissue Cells and serum-containing kidney cancer cells served as controls. Real-time and quantitative detection of telomerase activity in renal carcinoma stem cells was performed by telomeric repeat amplification (TRAP). Results: The expression rate of CD133 ~ + CD34 ~ + in serum-free suspension culture group was significantly higher than that in serum-containing culture group, the difference P <0.05 was statistically significant. Moreover, the telomerase activity of kidney cancer stem cells and kidney cancer cells were significantly higher than that of normal renal tissues, the difference P <0.05 was statistically significant. Conclusion: Compared with serum-free medium, the expression of CD133 + in renal cancer stem cells is higher than that in serum-free medium. However, the telomerase activity of renal cell carcinoma cells cultured in serum-free medium is higher than that in normal kidney tissue.