Isolation of cancer stem cells and the establishment of a H 2O 2-resistant cancer stem cell model

• Published in: Zhongguo xiu fu chong jian wai ke za zhi Vol. 33; no. 11; pp. 1433 - 1438
• Main Authors: Liu, Qingxi, Chen, Hongran, Li, Hui, Zhang, Tongcun, Ma, Wenjian
• Format: Journal Article
• Language: Chinese
• Published: 15.11.2019
• ISSN: 1002-1892
• DOI: 10.7507/1002-1892.201809014

OBJECTIVETo isolate cancer stem cells (CST) from human breast cancer cell line (MCF-7) and study their sensitivity toward oxidative stress. METHODSMCF-7 cells were cultured in serum-free suspension culture medium to identify cells forming the sphere phenotype. The morphological changes of MCF-7 cells were observed by inverted phase contrast microscope (compared with MCF-7 cells cultured in serum-free suspension culture medium). The expression of CST marker CD133 was detected by immunocytochemical staining in CST cell spheres (experimental group) with a diameter of 100 μm and MCF-7 cells (control group) with a fusion degree of 70%. The positive rate of CD133 was detected by flow cytometry in the third generation of tumor cells with diameter of 150 μm. The second generation of tumor globular cells (experimental group) with diameter of 150 μm and corresponding MCF-7 cells (control group) were taken to be damaged by 50 mol/L H 2O 2 for 120 minutes. The expression of DNA damage marker histone H2AX phosphorylation