Characterization of human fetal brain endothelial cells reveals barrier properties suitable for in vitro modeling of the BBB with syngenic co-cultures
Characterization of human fetal brain endothelial cells reveals barrier properties suitable for in vitro modeling of the BBB with syngenic co-cultures
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DOI：
10.1177/0271678x17708690
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发表时间：
2018-05-01
影响因子：
6.3
通讯作者：
Ramirez, Servio H.
中科院分区：
文献类型：
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作者：
Andrews, Allison M.;Lutton, Evan M.;Ramirez, Servio H.
内皮细胞（EC）形成血脑屏障（BBB）的基础，血脑屏障是一种选择性限制转运进入大脑的物理屏障。体外模型可以提供对BBB生理学、人类疾病病理学、毒理学和药物递送机制的重要见解。鉴于原代成人脑微血管内皮细胞（aBMVEC）的可用性有限，人胎儿组织为多个供体提供了一个合理的替代来源，并有机会从同一宿主中构建同基因三代培养物。以前培养胎儿脑微血管内皮细胞（fBMVECs）的努力没有成功地建立成熟的屏障特性。使用最佳胎龄进行分离和流式细胞术细胞分选，我们首次表明fBMVECs表现出成熟的屏障特性。当与aBMVEC相比时，fBMVEC表现出类似的功能表型，包括屏障完整性、内皮活化以及紧密连接蛋白和转运蛋白的基因/蛋白表达。重要的是，我们表明用于培养fBMVEC的组织也可用于产生同基因共培养物，在芯片上产生微流体BBB。提出的研究结果提供了一种手段，以克服以前的挑战，限制成功的屏障形成的fBMVECs。此外，该来源有利于神经血管单位的自体重建，用于下一代体外BBB建模。
Endothelial cells (ECs) form the basis of the blood-brain barrier (BBB), a physical barrier that selectively restricts transport into the brain. In vitro models can provide significant insight into BBB physiology, mechanisms of human disease pathology, toxicology, and drug delivery. Given the limited availability of primary human adult brain microvascular ECs (aBMVECs), human fetal tissue offers a plausible alternative source for multiple donors and the opportunity to build syngenic tri-cultures from the same host. Previous efforts to culture fetal brain microvascular ECs (fBMVECs) have not been successful in establishing mature barrier properties. Using optimal gestational age for isolation and flow cytometry cell sorting, we show for the first time that fBMVECs demonstrate mature barrier properties. fBMVECs exhibited similar functional phenotypes when compared to aBMVECs for barrier integrity, endothelial activation, and gene/protein expression of tight junction proteins and transporters. Importantly, we show that tissue used to culture fBMVECs can also be used to generate a syngenic co-culture, creating a microfluidic BBB on a chip. The findings presented provide a means to overcome previous challenges that limited successful barrier formation by fBMVECs. Furthermore, the source is advantageous for autologous reconstitution of the neurovascular unit for next generation in vitro BBB modeling.