1 Department of Thyroid and Breast Surgery, West China Hospital, Sichuan University, Chengdu 610041, China ; 2 Department of Breast Surgery, the Affiliated Hospital of Guiyang Medical College, Guiyang 550000, China ; 3 Laboratory of Stem Cell Biology, State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, China.
J Thorac Dis. 2014 Jun;6(6):829-37. doi: 10.3978/j.issn.2072-1439.2014.03.38.
Breast cancer stem cells (BCSCs) can be enriched by culturing of cells in non-adherent non-differentiating conditions. However, culturing mammospheres from primary breast tumors are costly and difficult to control. In order to overcome problems associated with using primary human tissues, continuous breast cancer cell lines have been developed from various sources.
In this study, a luminal subtype breast cancer cell line MCF-7 and a basal subtype cell line MDA-MB-231 were chosen. We explored the optimal culturing system for BCSCs from the two cell lines and primary breast tumors. Then, mammosphere formation efficiency (MFE), CD44(+)/CD24(-/low)ESA(+)Lin(-) cell proportion in mammospheres, and tumorigenecity of mammospheres generated from the two breast cancer cell lines and primary breast tumors were compared.
Enzymatic digestion of 60 mins and the addition of B27 to the culture medium were optimal for mammosphere culturing. Mammospheres could be formed in all the three cells, in which MCF-7 had the highest MFE. After 3 weeks culture, CD44(+)/CD24(-/low)ESA(+)Lin(-) cell proportion in mammospheres from MCF-7, MDA-MB-231 cells and primary breast tumors was 95.0%±2.5%, 82%±22% and 21.5%±1.0%, respectively. A total of 1,000 cells from MCF-7, MDA-MB-231 mammospheres but not primary mammospheres were tumorigenic.
This study validates the use of breast cancer cell lines as models to elucidate the nature of BCSCs.
乳腺癌干细胞（BCSCs）可通过在非贴壁非分化条件下培养细胞来富集。然而，从原发性乳腺癌培养类乳腺球体既昂贵又难以控制。为了克服使用原发性人组织相关的问题，已经从各种来源开发出了连续的乳腺癌细胞系。
在这项研究中，选择了腔型乳腺癌细胞系 MCF-7 和基底型细胞系 MDA-MB-231。我们探索了这两种细胞系和原发性乳腺癌的最佳 BCSC 培养系统。然后，比较了从两种乳腺癌细胞系和原发性乳腺癌生成的类乳腺球体的形成效率（MFE）、类乳腺球体中 CD44(+)/CD24(-/低)ESA(+)Lin(-)细胞的比例以及类乳腺球体的致瘤性。
60 分钟的酶消化和在培养基中添加 B27 是类乳腺球体培养的最佳条件。三种细胞均可以形成类乳腺球体，其中 MCF-7 的 MFE 最高。经过 3 周培养，MCF-7、MDA-MB-231 细胞和原发性乳腺癌来源的类乳腺球体中 CD44(+)/CD24(-/低)ESA(+)Lin(-)细胞的比例分别为 95.0%±2.5%、82%±22%和 21.5%±1.0%。只有 MCF-7 和 MDA-MB-231 的类乳腺球体中的 1000 个细胞而不是原发性类乳腺球体具有致瘤性。
这项研究验证了使用乳腺癌细胞系作为模型来阐明 BCSC 的性质。
