【摘要】 目的通过培养和富集乳腺癌细胞系及其干细胞,探讨不同乳腺癌细胞系中CD44+CD24-/low细胞比例,并分离鉴定乳腺癌干细胞。方法培养两种乳腺癌细胞系MCF-7、MDA-MB-231,利用流式细胞仪检测两种乳腺癌细胞系中CD44+CD24-/low细胞比例;通过无血清悬浮培养,观察其是否形成细胞球,再联合化疗筛选,富集乳腺癌干细胞。结果无血清悬浮培养,MDA-MB-231细胞未发现明显细胞球,MCF-7培养约6d出现细胞球;加入化疗药后,MCF-7细胞成球周期短至4d。流式细胞仪检测,MCF-7、MDA-MB-231两种细胞系中CD44+CD24-/low细胞比例分别2.07%和0.20%。结论利用CD44+CD24-/low标记流式细胞仪检测乳腺癌干细胞,简单有效;无血清悬浮培养,能富集乳腺癌干细胞,联合化疗双重筛选,能提高富集效率。更多还原

【Abstract】 Objective Through the cultivation and enrichment of breast cancer cell lines and their stem-cells,to discuss on different human breast cancer cell line CD44+CD24-/lowin proportion,and methods of isolation and identification of breast cancer stem-cells.Methods To train two breast cancer cell lines MCF-7,MDA-MB-231,using of flow cytometry detected CD44+CD24-/lowcells in two breast carcinoma cell lines;By serum-free suspension culturing,to observe whether they formed ball of cells,and then combined with chemotherapy to fliter and enrich breast cancer stem-cells.Results Serum-free suspension cultured,MDA-MB-231 ball of cells was not clear;and MCF-7ball cells was cultured about six days;After joining the chemotherapy drugs(TA Programme),MCF-7cell formed balls cycles as short as 4days.By flow cytometry,to detect the proportion of CD44+CD24-/low,MDA-MB-231 and MCF-7cell lines was 2.07% and 0.2% percent respectively.Conclusion Using the CD44+CD24-/lowtag to test breast cancer stem-cells by flow cytometry,it is simple and effective.Serum-free suspension culturing and combining with chemotherapy in a dual filter to enrich breast cancer stem-cells can improve the efficiency of accumulation.更多还原