Establishment of a Human Blood-Brain Barrier Co-culture Model Mimicking the Neurovascular Unit Using Induced Pluri- and Multipotent Stem Cells.
Establishment of a Human Blood-Brain Barrier Co-culture Model Mimicking the Neurovascular Unit Using Induced Pluri- and Multipotent Stem Cells.
复制标题
DOI：
10.1016/j.stemcr.2017.02.021
复制
发表时间：
2017-04-11
影响因子：
5.9
通讯作者：
Metzger M
中科院分区：
文献类型：
--
作者：
Appelt-Menzel A;Cubukova A;Günther K;Edenhofer F;Piontek J;Krause G;Stüber T;Walles H;Neuhaus W;Metzger M
人血脑屏障（BBB）的体外模型对于药物开发是非常理想的。本研究旨在分析基于原代细胞、人诱导多能干细胞（hiPSCs）和多能胎儿神经干细胞（fNSCs）的十种不同血脑屏障培养模型。我们系统地研究了星形胶质细胞、周细胞和NSCs对hipsc来源的血脑屏障内皮细胞功能和基因表达的影响。基于这四种细胞类型的四重培养模型实现了血脑屏障特征，包括高达2,500 Ω cm2的跨内皮电阻（TEER）和典型血脑屏障基因的明显上调。通过冷冻断裂和透射电镜检测到一个复杂的体内样紧密连接（TJ）网络。用claudin特异性TJ调节剂治疗导致TEER降低，证实了claudin亚型在细胞旁紧密性中的相关作用。用参比物质进行了药物渗透性试验，证实了模型对药物转运研究的适用性。基于hipsc和fNSCs的人血脑屏障标准化共培养模型的建立生理血脑屏障完整性和相关转运蛋白/TJs表达的反映通过clclin特异性TJ调节剂确认TJ网络功能通过通透性研究验证生理跨细胞模型的紧密性Metzger及其同事提出了基于诱导多能干细胞和多能干细胞的生理学相关的人类血脑屏障四重培养模型的建立。该模型可作为药物研究和ADMET研究的有力工具，使有希望的候选药物更可靠地转化为临床应用。
In vitro models of the human blood-brain barrier (BBB) are highly desirable for drug development. This study aims to analyze a set of ten different BBB culture models based on primary cells, human induced pluripotent stem cells (hiPSCs), and multipotent fetal neural stem cells (fNSCs). We systematically investigated the impact of astrocytes, pericytes, and NSCs on hiPSC-derived BBB endothelial cell function and gene expression. The quadruple culture models, based on these four cell types, achieved BBB characteristics including transendothelial electrical resistance (TEER) up to 2,500 Ω cm2 and distinct upregulation of typical BBB genes. A complex in vivo-like tight junction (TJ) network was detected by freeze-fracture and transmission electron microscopy. Treatment with claudin-specific TJ modulators caused TEER decrease, confirming the relevant role of claudin subtypes for paracellular tightness. Drug permeability tests with reference substances were performed and confirmed the suitability of the models for drug transport studies. Establishment of a standardized human BBB co-culture model based on hiPSCs and fNSCs Reflection of physiological BBB integrity and expression of relevant transporters/TJs Confirmation of TJ network functionality by claudin-specific TJ modulators Validation of physiological transcellular model tightness by permeability studies In this article, Metzger and colleagues present the establishment of physiologically relevant human blood-brain barrier quadruple culture models based on induced pluripotent and multipotent stem cells. The novel model can be used as a powerful tool in pharmaceutical drug research and ADMET studies, which may allow a more reliable translation of promising drug candidates into clinical application.