Pericytes augment the capillary barrier in in vitro cocultures
Pericytes augment the capillary barrier in in vitro cocultures
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DOI：
10.1006/jsre.2001.6117
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发表时间：
2001-05-01
影响因子：
2.2
通讯作者：
Tyburski, JG
中科院分区：
文献类型：
--
作者：
Dente, CJ;Steffes, CP;Tyburski, JG
大多数关于毛细血管通透性的体外研究都集中在内皮细胞(MVEC)单层，而忽略了形成毛细血管壁的第二个细胞：微血管周细胞(PC)。我们描述了一个模型来研究MVEC、PC和MVEC的通透性：PC共培养。半透性培养块包被胶原，然后与早期传代的牛肺微血管内皮细胞复合。在第3天加入牛肺PC，使其浓度接近MVEC：PC的1：1、5：1和10：1。在接下来的几天测量电阻，并在通透性分析中使用荧光标记(FITC)白蛋白来计算每个培养物的白蛋白清除量。电阻测量和白蛋白分析的结果也显示出类似的模式。内皮细胞单层阻力显著高于对照组，白蛋白通透性显著低于对照组。与单独内皮细胞相比，以10：1和5：1的比例添加周细胞可增加通透性屏障，尽管这些培养物彼此之间没有显著差异。1：1比例的混合培养具有最好的屏障，显著好于所有其他培养。内皮细胞单层是微循环的一个不充分的模型。由于PC在体内形成毛细血管壁的关键成分，以及在体外PC与MVEC单层的最佳比例为1：1，增加了它们对大小分子的屏障作用，我们认为在未来的体外研究中有必要将这两种细胞都包括在内。(C)2001年学术出版社。
Most in vitro studies of capillary permeability focus on endothelial cell (MVEC) monolayers and ignore the second cell that forms the capillary wall: the microvascular pericyte (PC). We describe a model to study the permeability characteristics of MVEC, PC, and MVEC:PC cocultures.Methods. Semipermeable culture inserts were coated with collagen and then plated with early passage bovine pulmonary MVEC. On Day 3, bovine pulmonary PC were added at concentrations to approximate MVEC:PC ratios of 1:1, 5:1, and 10:1. Electrical resistance was measured on subsequent days and fluorescently labeled (FITC) albumin was used in a permeability assay to calculate an albumin clearance for each culture.Results. The results for electrical resistance measurements and albumin assays showed a similar pattern. Resistance for endothelial cell monolayers was significantly higher and albumin permeability was significantly lower than that of controls. Addition of pericytes at a 10:1 and 5:1 ratios increased the permeability barrier compared to endothelial cells alone, although these cultures were not significantly different from one another. Cocultures at a 1:1 ratio had the best barrier, significantly better than all other cultures.Conclusions. Endothelial cell monolayers are an inadequate model of the microcirculation. As PC form a key component of the capillary wall in vivo and as addition of PC to MVEC monolayers in vitro, optimally at a 1:1 ratio, increase their barrier effect to large and small molecules, we believe it is necessary to include both cells in future in vitro studies. (C) 2001 Academic Press.