Human astrocytes/astrocyte-conditioned medium and shear stress enhance the barrier properties of human brain microvascular endothelial cells
Human astrocytes/astrocyte-conditioned medium and shear stress enhance the barrier properties of human brain microvascular endothelial cells
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DOI：
10.1016/j.brainres.2007.02.029
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发表时间：
2007-05-25
期刊：
影响因子：
2.9
通讯作者：
Kim, Kwang Sik
中科院分区：
文献类型：
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作者：
Siddharthan, Venkatraman;Kim, Yuri V.;Kim, Kwang Sik
血脑屏障(BBB)是一种结构和功能屏障，调节分子进出大脑以维持神经微环境。我们已经用人脑微血管内皮细胞(HBMEC)建立了体外血脑屏障模型。然而，在体内，HBMEC被证明与星形胶质细胞相互作用，并通过血液流动暴露于剪切力。为了建立模拟体内条件的血脑屏障模型，我们利用人脐静脉微血管内皮细胞和人胎儿星形胶质细胞(HFA)构建了体外血流基础血脑屏障模型。我们还研究了用星形胶质细胞条件培养液代替氢氟酸的效果，以研究分泌因子(S)对血脑屏障性质的影响。通过葡聚糖和碘化丙啶的通透性以及跨内皮细胞电阻(TEER)的测量来评价HBMEC单层的紧密性。结果表明，HFA和ACM非接触共培养可降低HBMEC的通透性，提高细胞的TEER。HBMEC暴露在剪应力下也表现出渗透性降低。此外，HFA/ACM和剪切流具有降低HBMEC单分子膜渗透性的相加作用。此外，我们还发现，与ACM共同培养并对切应力作出反应后，ZO-1(紧密连接蛋白)的HBMEC表达增加。这些结果表明，与HFA非接触共培养有助于维持HBMEC的屏障特性，其机制是通过分泌S因子来实现的。我们建立的人源性细胞体外流动模型系统可用于研究内皮细胞、神经胶质细胞和分泌因子(S)之间的相互作用以及剪切力在内皮细胞屏障性能中的作用。(C)2007 Elsevier B.V.保留所有权利。
The blood-brain barrier (BBB) is a structural and functional barrier that regulates the passage of molecules into and out of the brain to maintain the neural microenvironment. We have previously developed the in vitro BBB model with human brain microvascular endothelial cells (HBMEC). However, in vivo HBMEC are shown to interact with astrocytes and also exposed to shear stress through blood flow. In an attempt to develop the BBB model to mimic the in vivo condition we constructed the flow-based in vitro BBB model using HBMEC and human fetal astrocytes (HFA). We also examined the effect of astrocyte-conditioned medium (ACM) in lieu of HFA to study the role of secreted factor(s) on the BBB properties. The tightness of HBMEC monolayer was assessed by the permeability of dextran and propidium iodide as well as by measuring the transendothelial electrical resistance (TEER). We showed that the HBMEC permeability was reduced and TEER was increased by non-contact, co-cultivation with HFA and ACM. The exposure of HBMEC to shear stress also exhibited decreased permeability. Moreover, HFA/ACM and shear flow exhibited additive effect of decreasing the permeability of HBMEC monolayer. in addition, we showed that the HBMEC expression of ZO-1 (tight junction protein) was increased by co-cultivation with ACM and in response to shear stress. These findings suggest that the non-contact co-cultivation with HFA helps maintain the barrier properties of HBMEC by secreting factor(s) into the medium. our in vitro flow model system with the cells of human origin should be useful for studying the interactions between endothelial cells, glial cells, and secreted factor(s) as well as the role of shear stress in the barrier property of HBMEC. (c) 2007 Elsevier B.V. All rights reserved.