Aoki Hiromasa, Yamashita Misaki, Hashita Tadahiro, Iwao Takahiro, Aoyama Mineyoshi, Matsunaga Tamihide
Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya 467-8603, Japan.
Department of Pathobiology, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya 467-8603, Japan.
Pharmaceutics. 2022 Dec 2;14(12):2697. doi: 10.3390/pharmaceutics14122697.
Brain microvascular endothelial cells (BMECs) constitute the blood-brain barrier (BBB), which prevents the transfer of substances into the brain. Recently, in vitro BBB models using human-induced pluripotent stem (iPS) cell-derived brain microvascular endothelial-like cells (iBMELCs) have been created. However, it is suggested that iBMELCs differentiated by the existing methods are different from the BMECs that occur in vivo. This study aimed to establish iBMELCs generated via human iPS cell-derived endothelial progenitor cells (iEPCs) (E-iBMELCs). Expanded and cryopreserved iEPCs were thawed and differentiated into mature endothelial cells under various conditions. Intercellular barriers were significantly enhanced in E-iBMELCs using a B-27 supplement, transforming growth factor-β receptor inhibitor, and laminin 511 fragment. Expression of the endothelial cell markers was higher in the E-iBMELCs generated in this study compared with conventional methods. In addition, E-iBMELCs expressed P-glycoprotein. E-iBMELCs developed in this study will significantly contribute to drug discovery for neurodegenerative diseases and might elucidate the pathogenesis of neurodegenerative diseases associated with BBB disruption.
脑微血管内皮细胞（BMECs）构成血脑屏障（BBB），该屏障可阻止物质进入大脑。最近，利用人诱导多能干细胞（iPS）衍生的脑微血管内皮样细胞（iBMELCs）建立了体外血脑屏障模型。然而，有人提出，通过现有方法分化的iBMELCs与体内存在的BMECs不同。本研究旨在建立通过人iPS细胞衍生的内皮祖细胞（iEPCs）产生的iBMELCs（E-iBMELCs）。将扩增并冷冻保存的iEPCs解冻，并在各种条件下分化为成熟的内皮细胞。使用B-27补充剂、转化生长因子-β受体抑制剂和层粘连蛋白511片段，E-iBMELCs中的细胞间屏障显著增强。与传统方法相比，本研究中产生的E-iBMELCs中内皮细胞标志物的表达更高。此外，E-iBMELCs表达P-糖蛋白。本研究中开发的E-iBMELCs将对神经退行性疾病的药物发现做出重大贡献，并可能阐明与血脑屏障破坏相关的神经退行性疾病的发病机制。
