Derivation of Primary Choroid Plexus Epithelial Cells from the Mouse
Derivation of Primary Choroid Plexus Epithelial Cells from the Mouse
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DOI：
10.1007/978-1-59745-019-5_15
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发表时间：
2010-01-01
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影响因子：
--
通讯作者：
Ward, Andrew
中科院分区：
文献类型：
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作者：
Menheniott, Trevelyan R.;Charalambous, Marika;Ward, Andrew
脉络丛上皮细胞形成血液和脑脊液之间的屏障的不可分割的重要组成部分。脉络丛上皮的体外培养已经从几种哺乳动物物种中实现，这为脉络丛发育和功能的研究提供了机会，包括上皮细胞控制生物活性分子（如新的候选药物）从血流到脑的运动的能力。在这里，我们描述了一种方法的衍生原代细胞培养物从小鼠脉络丛上皮细胞，连同特征的免疫荧光标记的成熟脉络丛上皮细胞的特异性抗体。通过这种方法，使用DNA合成抑制剂阿糖胞苷（Ara-C）建立相对纯的脉络丛上皮细胞单层，所述DNA合成抑制剂阿糖胞苷（Ara-C）对污染细胞类型如成纤维细胞具有细胞毒性，但对上皮细胞没有细胞毒性。这些细胞显示表达诊断性脉络膜标志物，甲状腺素运载蛋白（TTR），以及上皮细胞分化的标志物，因此适合于研究，解决脉络丛的运输和屏障功能。
Choroid plexus epithelial cells form an integral and important part of the barrier between blood and cerebrospinal fluid. Culture of choroid plexus epithelium in vitro has been achieved from several mammalian species and this provides opportunities for the study of choroid plexus development and function, including the capacity of the epithelial cells to control the movement of bioactive molecules, such as novel drug candidates, from the bloodstream to the brain. Here we describe a method for the derivation of primary cell cultures from mouse choroid plexus epithelium, together with characterisation by immunofluorescence using antibodies specific to markers of mature choroid plexus epithelial cells. With this method, relatively pure choroid plexus epithelial cell monolayers are established using the DNA synthesis inhibitor cytosine arabinoside (Ara-C), which is cytotoxic to contaminating cell types such as fibroblasts, but not the epithelial cells. These cells are shown to express the diagnostic choroidal marker, transthyretin (TTR), as well as markers of epithelial cell differentiation and are thus suitable for studies that address the transport and barrier functions of the choroid plexus.