3D brain angiogenesis model to reconstitute functional human blood-brain barrier in vitro
3D brain angiogenesis model to reconstitute functional human blood-brain barrier in vitro
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DOI：
10.1002/bit.27224
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发表时间：
2019-12-04
影响因子：
3.8
通讯作者：
Jeon, Noo Li
中科院分区：
文献类型：
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作者：
Lee, Somin;Chung, Minhwan;Jeon, Noo Li
人类中枢神经系统（CNS）脉管系统表现出独特的屏障表型，即血脑屏障（BBB）。由于 BBB 通过限制药物转运导致中枢神经系统药物治疗效率低下，因此需要开发体外人类 BBB 模型。在这里，我们提出了中枢神经系统血管生成的微流体模型，其具有与血管周围细胞一致的三维（3D）腔化脉管系统。我们证实了血管生成三元培养系统（脑内皮与周细胞和星形胶质细胞直接相互作用）的必要性，以获得 BBB 脉管系统的基本表型，例如最小化的血管直径和最大化的连接表达。此外，与单一培养条件相比，在三重培养条件下实现了更低的血管通透性。值得注意的是，我们专注于重建功能性外排转运系统，包括 p-糖蛋白 (p-gp)，它对限制性药物转运负有重要责任。通过在外排转运蛋白抑制剂处理后对我们的3D可灌注脉管系统进行钙黄绿素-AM外排测定，我们证实了三培养模型中抑制剂具有更高的外排特性和显着效果。综上所述，我们基于受发育启发的 CNS 血管生成方案设计了具有功能屏障特性的 3D 人体 BBB 模型。我们期望该模型有助于更深入地了解病理性中枢神经系统血管生成和有效中枢神经系统药物的开发。
The human central nervous system (CNS) vasculature expresses a distinctive barrier phenotype, the blood-brain barrier (BBB). As the BBB contributes to low efficiency in CNS pharmacotherapy by restricting drug transport, the development of an in vitro human BBB model has been in demand. Here, we present a microfluidic model of CNS angiogenesis having three-dimensional (3D) lumenized vasculature in concert with perivascular cells. We confirmed the necessity of the angiogenic tri-culture system (brain endothelium in direct interaction with pericytes and astrocytes) to attain essential phenotypes of BBB vasculature, such as minimized vessel diameter and maximized junction expression. In addition, lower vascular permeability is achieved in the tri-culture condition compared to the monoculture condition. Notably, we focussed on reconstituting the functional efflux transporter system, including p-glycoprotein (p-gp), which is highly responsible for restrictive drug transport. By conducting the calcein-AM efflux assay on our 3D perfusable vasculature after treatment of efflux transporter inhibitors, we confirmed the higher efflux property and prominent effect of inhibitors in the tri-culture model. Taken together, we designed a 3D human BBB model with functional barrier properties based on a developmentally inspired CNS angiogenesis protocol. We expect the model to contribute to a deeper understanding of pathological CNS angiogenesis and the development of effective CNS medications.