Establishment and characterization of an immortalized Z310 choroidal epithelial cell line from murine choroid plexus
Establishment and characterization of an immortalized Z310 choroidal epithelial cell line from murine choroid plexus
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DOI：
10.1016/s0006-8993(02)03683-1
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发表时间：
2002-12-27
期刊：
影响因子：
2.9
通讯作者：
Zhao, QQ
中科院分区：
文献类型：
--
作者：
Zheng, W;Zhao, QQ
脉络丛在大脑发育中发挥着广泛的作用。成熟、衰老过程、内分泌调节和某些神经退行性疾病的发病机制。为了促进体外研究，我们使用基因转染技术使小鼠脉络膜上皮细胞永生化。通过磷酸钙沉淀将病毒质粒（pSV3neo）插入原代脉络膜上皮的宿主基因组中，从细胞毒性选择中存活下来的转染上皮细胞（即Z310细胞）在整个生命周期中表达SV40大T抗原，表明转染成功。通过光学显微镜观察，细胞显示出与起始细胞相同的多边形上皮形态。免疫细胞化学研究表明存在甲状腺素运载蛋白 (TTR)，这是一种已知仅由中枢神经系统脉络膜上皮产生的甲状腺素转运蛋白。在转染细胞和起始细胞中，蛋白质印迹分析进一步证实了这些细胞产生和分泌 TTR。通过 Northern 印迹分析鉴定编码转铁蛋白受体 (TfR) 的 mRNA。细胞以稳定的速度生长，目前处于第 110 代，在已建立的培养物中群体倍增时间为 20-22 小时。当Z310细胞在Trans-well装置上培养时，细胞形成类似于原代脉络膜细胞的上皮单层，具有内室和外室之间液位不均匀以及电阻约为150-200Ω-cm(2)等特征。这些结果表明永生化Z310细胞具有脉络膜上皮的特性，可能具有在血脑脊液屏障（BCB）研究中应用的潜力。 (C) 2002 Elsevier Science B.V. 保留所有权利。
The choroid plexus plays a wide range of roles in brain development. maturation, aging process, endocrine regulation, and pathogenesis of certain neurodegenerative diseases. To facilitate in vitro study, we have used a gene transfection technique to immortalize murine choroidal epithelial cells. A viral plasmid (pSV3neo) was inserted into the host genome of primary choroidal epithelia by calcium phosphate precipitation, The transfected epithelial cells, i.e., Z310 cells, that survived from cytotoxic selection expressed SV40 large-T antigen throughout the life span, suggesting a successful gone transfection. The cells displayed the same polygonal epithelial morphology as the starting cells by light microscopy. Immunocytochemical studies demonstrate the presence of transthyretin (TTR), a thyroxine transport protein known to be exclusively produced by the choroidal epithelia in the CNS. in both transfected and starting cells, Western blot analyses further confirm the production and secretion of TTR by these cells. The mRNAs encoding transferrin receptor (TfR) were identified by Northern blot analyses. The cells grow at a steady rate, currently in the 110th passage with a population doubling time of 20-22 h in the established culture. When Z310 cells were cultured onto a Trans-well apparatus, the cells formed an epithelial monolayer similar to primary choroidal cells, possessing features such as an uneven fluid level between inner and outer chambers and an electrical resistance approximately 150-200 Omega-cm(2). These results indicate that immortalized Z310 cells possess the characteristics of choroidal epithelia and may have the potential for application in blood-CSF barrier (BCB) research. (C) 2002 Elsevier Science B.V. All rights reserved.