无血清悬浮培养舌鳞癌Tca8113细胞系及肿瘤干细胞样亚群筛选

Serum-free suspension culture system and screening of stem like subpopulation in Tca8113 cell line of squamous cell carcinoma

目的:比较有血清培养基(Serum-supplemented medium,SSM)培养的细胞中和无血清培养基(Serum-free medium,SFM)培养的肿瘤干细胞球体中肿瘤干细胞样亚群的比例,以期获得较好富集肿瘤干细胞的方法。方法:SFM培养细胞,观察形成肿瘤干细胞球过程中的细胞形态;更换培养基为SSM诱导其分化;流式细胞仪检测SSM组细胞和SFM培养的肿瘤干细胞球分化前、后醛脱氢酶(Aldehyde dehydrogenase,ALDH)的表达。结果:Tca8113细胞能在SFM中存活、增殖并形成肿瘤干细胞球;SFM和SSM培养的细胞中ALDHhigh细胞比例分别为0.297 0±0.009 0和0.010 7±0.002 5,差异有统计学意义(P<0.05);诱导分化后细胞与SSM组细胞形态上无明显差异。结论:用SFM培养舌鳞癌Tca8113细胞可形成肿瘤干细胞球,而且这种细胞球富集了肿瘤干细胞。

Objective：To compare the proportion of stem like cells from serum-supplemented medium（SSM） culture with those from serum-free medium（SFM） culture in order to obtain better methods for enriching tumor stem cells.Methods： Tca8113 cells were cultivated in SFM.The cells were morphologically observed under optical microscope.To induce the differentiation of spherical cells,tumor spheres were cultivated in SSM.The expression of aldehyde dehydrogenase（ALDH） of undifferentiated spherical cel1s,post-differentiated spherical cells and SSM cells were detected by the fluorescence-activated cell sorting.Results：In SFM consisting of EGF,bFGF,and B27,a minority of Tca8113 cells could survive,proliferate,and form the suspended tumor spheres.The tumor spheres highly expressed ALDH（0.297 0±0.009 0∶0.010 7±0.002 5） and also displayed differentiation-induced abilities.No significant difference in morphology was found between the post-diferentiated spheres cells and routine cultured Tca8113 cells.Conclusion：Tumor spheres with enriched cancer stem cells can be generated under serum-free culture condition with EGF,bFGF,and B27.