IMMORTALIZATION AND FUNCTIONAL CHARACTERIZATION OF RAT ARACHNOID CELL LINES
IMMORTALIZATION AND FUNCTIONAL CHARACTERIZATION OF RAT ARACHNOID CELL LINES
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DOI：
10.1016/j.neuroscience.2010.12.035
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发表时间：
2011-03-17
期刊：
影响因子：
3.3
通讯作者：
Lam, C.
中科院分区：
文献类型：
--
作者：
Janson, C.;Romanova, L.;Lam, C.
对哺乳动物蛛网膜细胞的行为进行建模对于了解脑积水和其他涉及脑脊液异常流动的脑疾病至关重要，但由于缺乏强大的三维模型系统，对蛛网膜细胞的生理学了解相对较少。移植原代培养一直是研究蛛网膜细胞膜转运的唯一选择，但原代培养的实际局限性包括生长缓慢、早衰和重复性差。本研究的目的是建立永生化的大鼠蛛网膜细胞系，以便在体外研究蛛网膜颗粒和脑脊液(CSF)的流动特性。我们利用SV40大T抗原(SV40 LTAg)的逆转录病毒基因转移技术，建立了两个原代大鼠蛛网膜细胞永生化细胞系，并对其进行了部分鉴定。建立的细胞系稳定表达SV40 LTAg单独或SV40 LTAg和hTERT，并表现出高增殖率，融合时接触抑制，以及160多代天然蛛网膜细胞特有的蛋白质标志物的稳定表达。(C)2011年IBRO。爱思唯尔有限公司出版。保留所有权利。
Modeling the behavior of mammalian arachnoid cells is critical to understand hydrocephalus and other brain disorders involving abnormal flow of cerebrospinal fluid, yet relatively little is known about the physiology of arachnoid cells due to lack of a robust three-dimensional model system. Explanted primary cultures have been the only option to study transport across arachnoid cell membranes, but practical limitations of primary culture include slow growth, early senescence, and poor reproducibility. The purpose of this study was to create immortalized rat arachnoid cell lines to permit in vitro study of arachnoid granulations and properties of cerebrospinal fluid (CSF) flow. We established and partially characterized two immortalized cell lines generated from primary rat arachnoid cells, using retroviral gene transfer of SV40 large T antigen (SV40 LTAg) either with or without human telomerase (hTERT). The established cell lines stably express either SV40 LTAg alone, or SV40 LTAg and hTERT, and demonstrate high proliferative rate, contact inhibition at confluence, and stable expression of protein markers characteristic of native arachnoid cells over more than 160 passages. (C) 2011 IBRO. Published by Elsevier Ltd. All rights reserved.