无血清培养有效富集乳腺癌干细胞  

Enrichment of breast cancer stem cells by serum-free culture

作　　者：尹燕雪 李传伟[1] 于爱莲[2] 王学春[1] 

机构地区：[1]山东省泰山医学院病理学教研室,泰安271000 [2]山东省泰山医学院微生物学教研室,泰安271000

出　　处：《临床与实验病理学杂志》2015年第9期966-970,共5页Chinese Journal of Clinical and Experimental Pathology

基　　金：山东省科技攻关项目资助课题(2009GG20002047);山东省医药卫生科技发展计划(2011HD015)

摘　　要：目的探讨无血清培养技术从人乳腺癌MCF-7细胞株中有效富集乳腺癌干细胞的影响。方法采用无血清和有血清两种培养技术分别培养人乳腺癌MCF-7细胞株,观察并收集两种细胞,对CD24、CD44标志物进行免疫荧光染色;使用倒置荧光显微镜检测该标志物的表达差异;应用流式细胞仪检测不同亚群细胞的比例,并检测两种细胞周期的差异。结果无血清培养的细胞以大小不等的微球体形式在培养液中悬浮生长,有血清培养的细胞以类似多边形的形式在培养液中单层贴壁生长。CD44在悬浮微球体细胞和贴壁细胞中的表达无明显差异,CD24在悬浮微球体细胞中的表达明显减少;悬浮微球体细胞和贴壁细胞中CD44^+/ CD24^(-/low)表型细胞的比例分别为(86.93±0.53)%和(19.98±0.62)%(P<0.05),CD44^+/CD24^+表型细胞的比例分别为(12.68±0.59)%和(79.90±0.57)%(P<0.05)。分裂期(S)细胞在悬浮微球体细胞和贴壁细胞中的比例分别为(18.85±2.26)%和(43.91±1.81)%(P<0.05);静止期(G1)细胞的比例分别为(64.92±2.07)%和(39.82±1.77)%(P<0.05)。结论无血清培养技术可以有效富集CD44^+/ CD24^(-/low)表型乳腺癌干细胞。Purpose To identify whether serum-free culture can enrich breast cancer stem cells from MCF-7 human breast cancer cell line. Methods MCF-7 human breast cancer cell line by serum-free culture and serum culture technology were cultured, its cell mor-phology and growth pattern were observed by the inverted microscope. The expression of stem cell surface molecular makers CD24, CD44 was observed by the inverted fluorescence microscope and the flow cytometry, the proportion of different subpopulation cells was detected by the flow cytometry. At the same time, difference of the cell cycle was detected by the flow cytometry. Results The cell line cultured by serum-free culture grew in the form of suspended microspheres of different sizes in the medium, but the cell line cultured by serum culture grew in the form of monolayer adherent growth. There was no obvious difference in the expression of stem cell surface molecular makers CD44 between the suspended microsphere cells and the adherent cells, but the expression of CD24 in the sus-pended microsphere cells decreased compared to the adherent cells. The proportion of CD44^＋/CD24^-/low phenotype cells in the suspended microsphere cells and the adherent cells was （86. 93 ± 0. 53）% and （19. 98 ± 0. 62）%, respectively （P〈0. 05）, the proportion of CD44^ ＋ /CD24^ ＋ phenotype cells was （12. 68 ± 0. 59）% and （79. 90 ± 0. 57）%, respectively （P〈0. 05）. The proportion of mitotic cells in the suspended microsphere cells and the adherent cells was （ 18. 85 ± 2. 26 ）% and （ 43. 91 ± 1. 81 ）%, respectively （ P〈0. 05）, the proportion of quiescent cells was （64. 92 ± 2. 07）% and （39. 82 ± 1. 77）%, respectively （P〈0. 05）. Conclusion CD44^＋/CD24^-/low breast cancer stem cells can be effectively enriched by the serum-free culture technology.

关 键 词：乳腺肿瘤 干细胞 悬浮微球体细胞 CD44^+/CD24^-/low表型干细胞 

分 类 号：R737.9[医药卫生—肿瘤]