Program for Bioengineering, Seoul National University, Seoul, Korea.
Mechanical Engineering, Seoul National University, Seoul, Korea.
Biotechnol Bioeng. 2020 Mar;117(3):748-762. doi: 10.1002/bit.27224. Epub 2019 Dec 4.
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.
人类中枢神经系统 (CNS) 血管系统表达出独特的屏障表型，即血脑屏障 (BBB)。由于 BBB 通过限制药物转运来降低 CNS 药物治疗的效率，因此需要开发体外 BBB 模型。在这里，我们展示了一种具有三维 (3D) 管腔化血管的 CNS 血管生成的微流控模型，以及周细胞和星形胶质细胞的血管周围细胞。我们证实了血管生成三培养系统（脑内皮细胞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 血管的必要表型，例如最小化的血管直径和最大化的连接表达。此外，与单培养条件相比，三培养条件下的血管通透性更低。值得注意的是，我们专注于重建功能性外排转运体系统，包括对药物转运具有高度限制作用的 P-糖蛋白 (p-gp)。通过对我们的 3D 可灌注血管进行钙黄绿素 AM 外排测定，在用外排转运体抑制剂处理后，我们证实了三培养模型中更高的外排特性和抑制剂的显著作用。综上所述，我们根据 CNS 血管生成的发育启发式协议设计了具有功能性屏障特性的 3D 人 BBB 模型。我们期望该模型有助于深入了解病理性 CNS 血管生成，并开发有效的 CNS 药物。
