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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 extract breast cancer stem cells from MCF-7 breast cancer cells by cloning and identify them. Methods The breast cancer stem cells with “globular” growth were obtained by single cell clonal plus low density stem cell culture medium. The immunofluorescence cytochemical staining of CD44 and CD24 and differentiation induction experiments were performed. Results The proportion of CD44+CD24-/Low cells was 12.1%, and CD44+CD24-/Low cells were abundant in full cloning. The formed cell spheres are rich in CD44+CD24-/Low cells, and at the time of inducing differentiation, distinct luminal-like structures are formed, and CK18 and CK14 are expressed. Conclusion The cloning method is a simple and effective method for extracting cancer stem cells from cell lines.