Isolation of cancer stem-like cells from a side population of a human glioblastoma cell line, SK-MG-1
Isolation of cancer stem-like cells from a side population of a human glioblastoma cell line, SK-MG-1
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DOI：
10.1016/j.canlet.2009.10.010
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发表时间：
2010-05-28
期刊：
影响因子：
9.7
通讯作者：
Toda, Masahiro
中科院分区：
文献类型：
--
作者：
Fukaya, Raita;Ohta, Shigeki;Toda, Masahiro
越来越多的证据表明，在包括神经胶质瘤在内的几种类型的脑肿瘤中，只有一种称为脑癌干细胞（BCSC）的肿瘤细胞表型亚群可能能够启动肿瘤生长。最近，使用Hoechst染料分离侧群（SP）细胞已成为获得各种肿瘤中的癌症干细胞的有用方法。在这项研究中，我们使用SP技术从人脑胶质瘤细胞系中分离出肿瘤干细胞样细胞。流式细胞术分析显示，SK-MG-1，一种人胶质母细胞瘤细胞系，在所分析的五种胶质瘤细胞系中含有最多数量的SP细胞。SP细胞具有自我更新能力，并且能够在含有EGF和FGF 2的神经球培养基中形成球体。SP细胞的球体分化为神经元、星形胶质细胞和少突胶质细胞。RT-PCR分析显示SP细胞表达神经干细胞标记物Nestin。SP细胞在植入后8周在NOD/SCID小鼠的脑中产生肿瘤，而非SP细胞在脑中没有产生任何肿瘤。这些结果表明，从SK-MG-1分离的SP细胞具有癌症干细胞的特性，包括它们的自我更新能力、多谱系分化和致瘤性。因此，来自SK-MG-1的SP细胞可用于分析BCSC，因为它们易于处理和它们的产量。(C)2009爱思唯尔爱尔兰有限公司保留所有权利。
Accumulating evidence suggests that in several types of brain tumors, including glioma, only a phenotypic subset of tumor cells called brain cancer stem cells (BCSCs) may be capable of initiating tumor growth. Recently, the isolation of side population (SP) cells using Hoechst dye has become a useful method for obtaining cancer stem cells in various tumors. In this study, we isolated cancer stem-like cells from human glioma cell lines using the SP technique. Flow cytometry analysis revealed that SK-MG-1, a human glioblastoma cell line, contained the largest number of SP cells among the five glioma cell lines that were analyzed. The SP cells had a self-renewal ability and were capable of forming spheres in a neurosphere culture medium containing EGF and FGF2. Spheres derived from the SP cells differentiated into three different lineage cells: neurons, astrocytes and oligodendrocytes. RT-PCR analysis revealed that the SP cells expressed a neural stem cell marker, Nestin. The SP cells generated tumors in the brains of NOD/SCID mice at 8 weeks after implantation, whereas the non-SP cells did not generate any tumors in the brain. These results indicate that SP cells isolated from SK-MG-1 possess the properties of cancer stem cells, including their self-renewal ability, multi-lineage differentiation, and tumorigenicity. Therefore, the SP cells from SK-MG-1 may be useful for analyzing BCSCs because of the ease with which they can be handled and their yield. (C) 2009 Elsevier Ireland Ltd. All rights reserved.