Pericytes from brain microvessels strengthen the barrier integrity in primary cultures of rat brain endothelial cells
Pericytes from brain microvessels strengthen the barrier integrity in primary cultures of rat brain endothelial cells
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DOI：
10.1007/s10571-007-9195-4
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发表时间：
2007-09-01
影响因子：
4
通讯作者：
Niwa, Masami
中科院分区：
文献类型：
--
作者：
Nakagawa, Shinsuke;Deli, Maria A.;Niwa, Masami
(1)脑内皮细胞的血脑屏障（BBB）特性是由器官特异性局部信号诱导的。脑内皮细胞在没有与邻近细胞串扰的培养物中失去其表型。(2)与星形胶质细胞相反，周细胞（脑毛细血管中内皮细胞的另一邻近细胞）很少用于BBB共培养系统。(3)在培养插入物上由原代大鼠脑内皮细胞、星形胶质细胞和周细胞构建了七种不同类型的BBB模型，即单培养、双培养和三重共培养。通过测量跨内皮电阻和小分子量标记物荧光素的渗透性来比较模型的屏障完整性。(4)我们可以证实，在单一培养的脑内皮细胞单层不形成紧密的屏障。在共培养条件下，周细胞比I型星形胶质细胞诱导更高的电阻和更低的荧光素渗透性。在三重共培养模型中，当内皮细胞和周细胞位于插入物的多孔滤膜的两侧并且星形胶质细胞位于培养皿底部时，观察到最紧密的屏障。(5)首次建立了以原代培养为基础的大鼠同系三重共培养血脑屏障模型，该模型除脑内皮细胞和星形胶质细胞外，还使用了脑周细胞。该模型，密切模仿的解剖位置的细胞在血脑屏障在体内，是优于其他血脑屏障模型测试上级。(6)周细胞对脑内皮细胞BBB特性的影响可能与星形胶质细胞的影响一样重要，并可用于构建更好的BBB模型。
(1) The blood-brain barrier (BBB) characteristics of cerebral endothelial cells are induced by organ-specific local signals. Brain endothelial cells lose their phenotype in cultures without cross-talk with neighboring cells. (2) In contrast to astrocytes, pericytes, another neighboring cell of endothelial cells in brain capillaries, are rarely used in BBB co-culture systems. (3) Seven different types of BBB models, mono-culture, double and triple co-cultures, were constructed from primary rat brain endothelial cells, astrocytes and pericytes on culture inserts. The barrier integrity of the models were compared by measurement of transendothelial electrical resistance and permeability for the small molecular weight marker fluorescein. (4) We could confirm that brain endothelial monolayers in mono-culture do not form tight barrier. Pericytes induced higher electrical resistance and lower permeability for fluorescein than type I astrocytes in co-culture conditions. In triple co-culture models the tightest barrier was observed when endothelial cells and pericytes were positioned on the two sides of the porous filter membrane of the inserts and astrocytes at the bottom of the culture dish. (5) For the first time a rat primary culture based syngeneic triple co-culture BBB model has been constructed using brain pericytes beside brain endothelial cells and astrocytes. This model, mimicking closely the anatomical position of the cells at the BBB in vivo, was superior to the other BBB models tested. (6) The influence of pericytes on the BBB properties of brain endothelial cells may be as important as that of astrocytes and could be exploited in the construction of better BBB models.