目的

通过克隆柱法提取MCF-7乳腺癌细胞中的乳腺癌干细胞,并进行鉴定。

方法

采用单细胞克隆加低密度干细胞培养基筛选获得呈“球样”生长的乳腺癌干细胞,行CD44、CD24免疫荧光细胞化学染色及诱导分化实验。

结果

CD44⁺CD24^-/Low细胞的比例为12.1%,且在全克隆中富含CD44⁺CD24^-/Low 细胞。在形成的细胞球中富含CD44⁺CD24^-/Low细胞,并且在诱导分化时可见明显类管腔样结构形成,并表达CK18及CK14。

结论

克隆柱法是提取细胞系中癌干细胞的简单有效方法。

Objective

To isolate cancer stem-like cells from MCF-7 breast cancer cell line with modified cloning cylinder method and to test its feasibility.

Methods

Single cell cloning and stem cell screening with stem cell culture medium were applied to obtain sphere-like breast cancer stem cells.Immunofluorescence staining with CD44 and CD24 and inducement and differentiation experiment were performed.

Results

The proportion of CD44⁺CD24^-/Low cells was 12.1%,and the proportion in the holoclone was high.The majority of cells in culturing mammospheres were stained positively for CD44 and negatively for CD24.The mammospheres had abundant CD44⁺CD24^-/Low cells.Sphere-like breast cancer stem cells were obtained from holoclone cells,which were seeded at low density and suspension cultured with stem cell culture medium.When the sphere-like breast cancer stem cells were induced to differentiate,duct-like structures emerged and expressed CK18 and CK14.

Conclusion

The cloning cylinder method is a simple and effective way to obtain breast cancer stem celllike cells from MCF-7 breast cancer cell lines.