无血清悬浮法培养MCF-7细胞系并筛选该细胞系中肿瘤干细胞相关亚群的初步研究

被引：16

作者：

许立生 ^{[1
]}

王水 ^{[1
]}

许健 ^{[1
]}

杜青 ^{[2
]}

机构：

[1] 南京医科大学第一附属医院乳腺外科

[2] 南京医科大学第一附属医院普外科实验室

来源：

中华肿瘤防治杂志 | 2007年 / 10期

关键词：

乳腺肿瘤/病理学; 肿瘤干细胞/病理学; 细胞系,肿瘤; 细胞培养技术/方法;

D O I：

10.16073/j.cnki.cjcpt.2007.10.005

中图分类号：

R737.9 [乳腺肿瘤];

学科分类号：

摘要：

目的:研究无血清培养基悬浮培养MCF-7乳腺癌细胞系,筛选并鉴定MCF-7乳腺癌细胞系中的肿瘤干细胞相关亚群。方法:应用乳腺癌培养基在无血清的条件下悬浮培养MCF-7乳腺癌细胞系。通过无血清培养筛选乳腺癌细胞系肿瘤干细胞相关亚群,将其接种于含血清培养基,观察分化。应用单克隆形成实验、表面标志检测、HOECHST33342染色检测来确定培养出的细胞中肿瘤干细胞的比例及其培养后肿瘤干细胞含量的变化。结果:乳腺癌MCF-7细胞系中有约2.12%的肿瘤细胞在无血清培养基中能够存活、增殖,形成自由漂浮的细胞球。细胞球可连续传代,若重新接种于含血清培养基中可重新贴壁分化,贴壁分化后细胞形态与直接在含血清培养基中培养的MCF-7无明显差别。流式细胞仪表面标志检测,细胞球中约含83.13%表达CD24-CD44+的细胞,HOECHST33342染色提示细胞球中约含10.06%的侧亚群(sidepopulation)细胞。结论:MCF-7细胞系可在含生长因子的无血清培养基中悬浮生长,并维持细胞系。该细胞系中含有比例较低的具有增殖和分化能力的乳腺癌干细胞相关亚群。表面标志以及HOECHST33342检测差异提示细胞球中仅约1/8细胞具有干细胞的功能。

引用

页码：736 / 740

页数：5

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