iPSC-Derived Brain Endothelium Exhibits Stable, Long-Term Barrier Function in Perfused Hydrogel Scaffolds
iPSC-Derived Brain Endothelium Exhibits Stable, Long-Term Barrier Function in Perfused Hydrogel Scaffolds
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DOI：
10.1016/j.stemcr.2019.01.009
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发表时间：
2019-03-05
影响因子：
5.9
通讯作者：
Bellan, Leon M.
中科院分区：
文献类型：
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作者：
Faley, Shannon L.;Neal, Emma H.;Bellan, Leon M.
对人血脑屏障和神经血管单元（NVU）的功能性仿生体外组织构建体存在深刻的需求，以模拟疾病并鉴定治疗干预。在这里，我们表明，诱导多能干细胞（iPSC）衍生的人脑微血管内皮细胞（BMEC）在明胶水凝胶内的3D通道中培养时表现出强大的屏障功能。我们确定，在灌注条件下在3D中培养的BMEC相对于人脐静脉内皮细胞和人真皮微血管内皮细胞对照对荧光素钠、3 kDa葡聚糖和白蛋白的渗透性低10-100倍，并且BMEC维持屏障功能长达21天。细胞-细胞连接的分析揭示了支持屏障形成的表达模式。最后，外排转运蛋白的活性维持了3周的灌注培养。总之，这项工作为开发由iPSC构建的人NVU的代表性3D体外模型奠定了基础。
There is a profound need for functional, biomimetic in vitro tissue constructs of the human blood-brain barrier and neurovascular unit (NVU) to model diseases and identify therapeutic interventions. Here, we show that induced pluripotent stem cell (iPSC)-derived human brain microvascular endothelial cells (BMECs) exhibit robust barrier functionality when cultured in 3D channels within gelatin hydrogels. We determined that BMECs cultured in 3D under perfusion conditions were 10-100 times less permeable to sodium fluorescein, 3 kDa dextran, and albumin relative to human umbilical vein endothelial cell and human dermal microvascular endothelial cell controls, and the BMECs maintained barrier function for up to 21 days. Analysis of cell-cell junctions revealed expression patterns supporting barrier formation. Finally, efflux transporter activity was maintained over 3 weeks of perfused culture. Taken together, this work lays the foundation for development of a representative 3D in vitro model of the human NVU constructed from iPSCs.