肾癌干细胞的培养鉴定及其凋亡的实验研究

Isolation and identification of RCSC and study of its apoptosis property

目的探讨肾癌干细胞在肾癌的发生发展中的作用。方法收集肾癌细胞检测其CD133表达情况,并研究其凋亡特性;然后将其悬浮于含有表皮生长因子(EGF)和成纤维生长因子(bFGF)的无血清培养基中培养7d,收集干细胞球,同时收集利用含10%胎牛血清培养基培养的肾癌细胞作阴性对照,分别检测其CD133表达情况,并研究其凋亡特性。结果悬浮培养2d后出现肾癌干细胞球,培养7d后干细胞球明显增多,收集干细胞球,用流式细胞仪检测发现表达CD133+细胞约占(8.25±1.43)%,阴性对照中CD133+细胞只占(1.22±0.34)%,并且较前者具有较高的凋亡率。结论利用无血清体外培养并收集悬浮生长的肾癌细胞的方法可以从肾癌细胞中获取肾癌干细胞;肾癌干细胞较一般瘤组织有较低的凋亡率,维持着肿瘤的无限增殖。

Objective To enrich renal cell carcinoma（RCC）stem cells from RCC cells line in serum free medium（SFM）with the method of suspension culture and to study its apoptosis.Methods RCC cells were collected and dissotiated into single ones whose expression of CD133 and apoptotic rate were first detected,then they were seeded in SFM containing EGF and bFGF.Seven days later the expression of CD133 and apoptotic rate in cells from SFM were detected by flow cytometry and normal RCC cells were set as negative control.Results RCC stem cells spheres were appeared after RCC cells were cultured tow days in SFM ;Seven days later,more spheres came out.Then collected and detected them by flow cytometry and found that about（1.22±0.34）% cells were CD133＋cells in normal OS-RC-2 cell line,while CD133＋cells reached to（8.25±1.43）% after cultivating in SFM for 7 days and had low apoptotic rate.Conclusion RCC stem cells can be enriched from RCC cells by SFM,and they have lower apoptotic rate than normal RCC cells...更多 and are keeping their unlimited proliferating.