Monnot Andrew D, Zheng Wei
School of Health Sciences, Purdue University, West Lafayette, IN, USA.
Methods Mol Biol. 2013;945:13-29. doi: 10.1007/978-1-62703-125-7_2.
Chemical homeostasis in the extracellular fluid of the central nervous system (CNS) is maintained by two brain barrier systems, i.e., the blood-brain barrier (BBB) that separates the blood circulation from brain interstitial fluid and the blood-cerebrospinal fluid barrier (BCB) that separates the blood from the cerebrospinal fluid (CSF). The choroid plexus, where the BCB is located, is a polarized tissue, with the basolateral side of the choroidal epithelium facing the blood and the apical microvilli in direct contact with the CSF. The tissue plays a wide range of roles in brain development, aging, nutrient transport, endocrine regulation, and pathogenesis of certain neurodegenerative disorders. This chapter describes two in vitro cultures that have been well established to allow for study of the BCB structure and function. The primary choroidal epithelial cell culture can be established from rat choroid plexus tissue, and a similar immortalized murine choroidal epithelial cell culture known as Z310 cells has also been established. Both cultures display a dominant polygonal morphology, and immunochemical studies demonstrate the presence of transthyretin, a thyroxine transport protein known to be exclusively produced by the choroidal epithelia in the CNS. These cultures have been adapted for use on freely permeable Transwell(®) membranes sandwiched between two culture chambers, facilitating transport studies of various compounds across this barrier in vitro. These choroidal epithelia cultures with the Transwell system will perceivably assist blood-CSF barrier research.
中枢神经系统(CNS)细胞外液中的化学稳态由两个脑屏障系统维持,即血脑屏障(BBB),它将血液循环与脑间质液分隔开;以及血脑脊液屏障(BCB),它将血液与脑脊液(CSF)分隔开。血脑脊液屏障所在的脉络丛是一种极化组织,脉络丛上皮的基底外侧面对着血液,顶端微绒毛与脑脊液直接接触。该组织在脑发育、衰老、营养物质运输、内分泌调节以及某些神经退行性疾病的发病机制中发挥着广泛作用。本章描述了两种已被充分建立的体外培养方法,用于研究血脑脊液屏障的结构和功能。原代脉络丛上皮细胞培养可以从大鼠脉络丛组织中建立,并且还建立了一种类似的永生化小鼠脉络丛上皮细胞培养物,称为Z310细胞。两种培养物都呈现出占主导的多边形形态,免疫化学研究表明存在转甲状腺素蛋白,这是一种已知仅由中枢神经系统中的脉络丛上皮产生的甲状腺素转运蛋白。这些培养物已被适配用于夹在两个培养室之间的可自由渗透的Transwell®膜上,便于体外研究各种化合物跨此屏障的转运。这些带有Transwell系统的脉络丛上皮细胞培养物有望辅助血脑脊液屏障的研究。