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目的从人卵巢癌细胞系A2780中分离并鉴定卵巢癌干细胞。方法将A2780细胞置于含人重组表皮生长因子(epidermal growth factor,EGF)、人重组碱性成纤维生长因子(basic fibroblast growth factor,bFGF)、牛血清白蛋白和人胰岛素的无血清培养基中培养,采用连续传代诱导培养肿瘤干细胞,对每代分离出来的肿瘤干细胞进行流式检测,并对第4代肿瘤干细胞进行免疫荧光检测和逆向分化实验。结果从A2780细胞中成功分离出肿瘤干细胞,在上述无血清培养基中呈悬浮生长,具有很强的自我更新能力,流式检测显示随着传代诱导,CD133阳性率依次升高,免疫荧光进一步证实了CD133是卵巢癌干细胞的表面标记物,诱导分化实验说明其可逆向分化成为A2780肿瘤细胞。结论体外培养的卵巢癌细胞株A2780中存在卵巢癌干细胞,并能将其分离培养和诱导分化。
Objective To isolate and identify ovarian cancer stem cells from human ovarian cancer cell line A2780. Methods A2780 cells were plated in serum-free medium containing human recombinant epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), bovine serum albumin and human insulin The cultured stem cells were cultured and induced by continuous passage. The tumor stem cells isolated from each generation were tested by flow cytometry. The 4th passage cancer stem cells were detected by immunofluorescence and reverse differentiation. Results Tumor stem cells were successfully isolated from A2780 cells and grew in suspension in the above serum-free medium with high self-renewal ability. Flow cytometry showed that the positive rate of CD133 increased with the passage of time, and the immunofluorescence further confirmed CD133 is a surface marker of ovarian cancer stem cells, inducing differentiation experiments show that it can reverse differentiation into A2780 tumor cells. Conclusion Ovarian cancer stem cells exist in ovarian cancer cell line A2780 cultured in vitro and can be isolated, cultured and induced to differentiate.