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大鼠脉络丛原代上皮细胞培养方法

Method for Primary Epithelial Cell Culture from the Rat Choroid Plexus.

作者信息

Lallai Valeria, Ahmed Amina, Fowler Christie D

机构信息

Department of Neurobiology and Behavior, University of California Irvine, Irvine, CA, USA.

出版信息

Bio Protoc. 2020 Feb 20;10(4):e3532. doi: 10.21769/BioProtoc.3532.

Abstract

The choroid plexus consists of a network of secretory epithelial cells localized throughout the lateral, third and fourth ventricles of the brain. Cerebrospinal fluid (CSF) is generated by the choroid plexus and released into the ventricular environment. This biofluid contains an enriched source of proteins, ions, and other signaling molecules for extracellular support of neurons and glial cells within the central nervous system. Given that other cells in the brain also release factors into the CSF, investigations of choroid plexus function are necessary to isolate processes selectively occurring within and released from this tissue. Here, we describe a protocol to isolate choroid plexus tissue from each of the ventricular locations, and the cell culture conditions required to support growth and maintenance of these epithelial cells. This technique allows for investigations of the functional significance of the choroid plexus, such as for the examination of stimuli promoting the release of growth factors and extracellular vesicles , exosomes and microvesicles) from ventricle-specific choroid plexus epithelial cells.

摘要

脉络丛由分布于脑的侧脑室、第三脑室和第四脑室的分泌上皮细胞网络组成。脑脊液(CSF)由脉络丛产生并释放到脑室环境中。这种生物流体含有丰富的蛋白质、离子和其他信号分子来源,用于中枢神经系统内神经元和神经胶质细胞的细胞外支持。鉴于脑内的其他细胞也会向脑脊液中释放因子,因此有必要对脉络丛功能进行研究,以分离在该组织内选择性发生和从该组织释放的过程。在这里,我们描述了一种从每个脑室位置分离脉络丛组织的方案,以及支持这些上皮细胞生长和维持所需的细胞培养条件。这项技术有助于研究脉络丛的功能意义,例如检查促进特定脑室脉络丛上皮细胞释放生长因子和细胞外囊泡(外泌体和微囊泡)的刺激因素。

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