Blood-brain barrier-specific properties of a human adult brain endothelial cell line
Blood-brain barrier-specific properties of a human adult brain endothelial cell line
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DOI：
10.1096/fj.04-3458fje
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发表时间：
2005-09-01
期刊：
影响因子：
4.8
通讯作者：
Couraud, PO
中科院分区：
文献类型：
--
作者：
Weksler, BB;Subileau, EA;Couraud, PO
事实证明，建立人体血脑屏障模型是一个困难的目标。为了实现这一目标，通过掺入人端粒酶或 SV40 T 抗原的慢病毒载体转导正常人脑内皮细胞。在通过连续有限稀释克隆转导细胞获得的许多稳定的永生化克隆中，选择了一个表达正常内皮标志物，包括CD31、VE钙粘蛋白和血管性血友病因子。该细胞系称为 hCMEC/D3，显示出稳定的正常核型，在组织培养中维持接触抑制的单层，表现出响应内皮生长因子的强劲增殖，并在基质中形成毛细管，但在软琼脂中没有集落。 hCMEC/D3 细胞表达端粒酶并无限生长，无表型去分化。这些细胞表达趋化因子受体，响应炎症细胞因子上调粘附分子，并表现出血脑屏障特征，包括紧密连接蛋白和主动排除药物的能力。 hCMEC/D3 是研究血脑屏障功能、脑内皮对炎症和感染刺激的反应以及脑内皮与淋巴细胞或肿瘤细胞相互作用的优秀候选者。因此，hCMEC/D3代表了第一个稳定的、完全表征的、分化良好的人脑内皮细胞系，并且应该作为一种广泛使用的研究工具。
Establishment of a human model of the blood-brain barrier has proven to be a difficult goal. To accomplish this, normal human brain endothelial cells were transduced by lentiviral vectors incorporating human telomerase or SV40 T antigen. Among the many stable immortalized clones obtained by sequential limiting dilution cloning of the transduced cells, one was selected for expression of normal endothelial markers, including CD31, VE cadherin, and von Willebrand factor. This cell line, termed hCMEC/D3, showed a stable normal karyotype, maintained contact-inhibited monolayers in tissue culture, exhibited robust proliferation in response to endothelial growth factors, and formed capillary tubes in matrix but no colonies in soft agar. hCMEC/D3 cells expressed telomerase and grew indefinitely without phenotypic dedifferentiation. These cells expressed chemokine receptors, up-regulated adhesion molecules in response to inflammatory cytokines, and demonstrated blood-brain barrier characteristics, including tight junctional proteins and the capacity to actively exclude drugs. hCMEC/D3 are excellent candidates for studies of blood-brain barrier function, the responses of brain endothelium to inflammatory and infectious stimuli, and the interaction of brain endothelium with lymphocytes or tumor cells. Thus, hCMEC/D3 represents the first stable, fully characterized, well-differentiated human brain endothelial cell line and should serve as a widely usable research tool.