摘要
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 ovarian cancer cell line A2780.Methods The A2780 cells were cultured in serum-free medium with EGF,bFGF,insulin and BSA to induce tumor stem cells by continuous passage.Flow cytometry was applied to identify the cancer stem cells in every passage.Immunofluorescence staining and reverse differentiation experiments were conducted on the fourth passage of cancer stem cells.Results The ovarian cancer stem cells were successfully isolated from ovarian cancer cell line A2780.They formed typical neurospheres in serum-free medium,and had strong self-renew capacity.The results of flow cytometry illustrated that CD133 positive rate was increased with the passages of cancer stem cells.Immunofluorescence staining further confirmed that CD133＋ cells were the surface marker of ovarian cancer stem cells,and they could differentiate into ovarian cancer cells in reverse differentiation experiments.Conclusion There were ovarian cancer stem cells in ovarian cancer cell line A2780,which can be isolated,cultured and differentiated.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第4期356-359,共4页
Journal of Third Military Medical University
基金
国家自然科学基金(30901592)~~
关键词
卵巢肿瘤
肿瘤细胞
培养的
肿瘤干细胞
分离
鉴定
ovarian neoplasms
tumor cells
cultured
neoplastic stem cells
isolation
identification
