Menheniott Trevelyan R, Charalambous Marika, Ward Andrew
Murdoch Children's Research Institute, Royal Children's Hospital, Parkville, VIC 3052, Australia.
Methods Mol Biol. 2010;633:207-20. doi: 10.1007/978-1-59745-019-5_15.
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.
脉络丛上皮细胞构成了血液与脑脊液之间屏障的一个不可或缺且重要的部分。已成功从多种哺乳动物中实现脉络丛上皮细胞的体外培养，这为研究脉络丛的发育和功能提供了机会，包括上皮细胞控制生物活性分子（如新型候选药物）从血液进入大脑的移动能力。在此，我们描述一种从小鼠脉络丛上皮细胞获得原代细胞培养物的方法，以及使用针对成熟脉络丛上皮细胞标志物的特异性抗体通过免疫荧光进行表征的方法。使用这种方法，利用对成纤维细胞等污染细胞类型具有细胞毒性但对上皮细胞无毒性的DNA合成抑制剂阿糖胞苷（Ara-C）建立了相对纯净的脉络丛上皮细胞单层。这些细胞显示表达诊断性脉络丛标志物甲状腺转运蛋白（TTR）以及上皮细胞分化标志物，因此适用于研究脉络丛的转运和屏障功能。
