An efficient method for derivation and propagation of glioblastoma cell lines that conserves the molecular profile of their original tumours
An efficient method for derivation and propagation of glioblastoma cell lines that conserves the molecular profile of their original tumours
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DOI：
10.1016/j.jneumeth.2008.07.022
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发表时间：
2009-01-30
影响因子：
3
通讯作者：
Watts, Colin
中科院分区：
文献类型：
--
作者：
Al-Mayhani, Talal M. Fael;Ball, Siolian L. R.;Watts, Colin
越来越多的证据表明，胶质瘤干细胞样细胞更代表他们的父母肿瘤时，培养在定义的无血清条件下与有丝分裂原表皮生长因子（EGF）和成纤维细胞生长因子（FGF）。然而，将这些细胞培养为自由漂浮的球状体可能导致难以有效地衍生和繁殖细胞系。我们结合了神经球和单层培养技术，以提高效率与细胞可以从临床肿瘤样品中获得在定义的无血清conditions.We应用我们的协议连续样本的胶质母细胞瘤，以表明他们可以形成实验肿瘤，概括了许多的组织学特征的父母turnout。我们继续表明，turnout起始细胞也保留了亲本turnout的细胞遗传学异常。最后，我们检测了细胞系中与神经干细胞相关的标志物的表达。我们的研究结果证实了与神经模式和规范相关的转录因子的表达，包括Sox 2，Olig2，Pax 6和Nkx 2.2。我们继续建立，这些因素也表示在父母turnout表明，他们的表达是不是我们的培养condition.The剑桥协议的功能是一个有效的方法，从胶质母细胞瘤干细胞样肿瘤起始细胞。提高衍生化效率将有助于改进体外和体内模型系统，以研究疾病机制，筛选药物和开发新的治疗方法。(C)2008 Elsevier B.V.保留所有权利。
A growing body of evidence suggests that glioma stem-like cells are more representative of their parent tumours when cultured under defined serum-free conditions with the mitogens epidermal growth factor (EGF) and fibroblast growth factor (FGF). However, culturing these cells as free-floating spheroids can result in difficulty in efficiently deriving and propagating cell lines. We have combined neurosphere and monolayer culture techniques to improve the efficiency with which cells can be derived from clinical tumour samples under defined serum-free conditions.We have applied our protocol to consecutive samples of glioblastoma to show that they can form experimental tumours that recapitulate many of the histological features of the parent turnout. We go on to show that the turnout initiating cells also retain the cytogenetic abnormalities of the parent turnout.Finally we examined the cell lines for expression of markers associated with neural stem cells. Our results confirm the expression of transcription factors associated with neural patterning and specification including Sox2, Olig2, Pax6 and Nkx2.2. We went on to establish that these factors were also expressed in the parent turnout indicating that their expression was not a function of our culture conditions.The Cambridge Protocol is an efficient method of deriving stem-like tumour initiating cells from glioblastoma. improving the efficiency of derivation will facilitate the improvement of in vitro and in vivo model systems to study disease mechanisms, screen drugs and develop novel therapeutic approaches in the future. (C) 2008 Elsevier B.V. All rights reserved.