Tissue-engineered blood-brain barrier models via directed differentiation of human induced pluripotent stem cells
Tissue-engineered blood-brain barrier models via directed differentiation of human induced pluripotent stem cells
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DOI：
10.1038/s41598-019-50193-1
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发表时间：
2019-09-27
影响因子：
4.6
通讯作者：
Searson, Peter C.
中科院分区：
文献类型：
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作者：
Grifno, Gabrielle N.;Farrell, Alanna M.;Searson, Peter C.
血脑屏障(BBB)的三维(3D)组织工程模型概括了体内剪切力、圆柱形几何形状和细胞-细胞外基质相互作用。在这里，我们解决了与血脑屏障模型相关的四个问题：细胞源、屏障功能、冷冻保存和基质硬度。我们从两个荧光标记的人诱导多能干细胞系(HiPSCs)中复制了脑微血管内皮细胞(DhBMECs)的定向分化，并证明了荧光黄在六天内的生理渗透性。从深低温保存的dhBMEC形成的微血管显示出BBB标志的表达，并保持与未深低温保存的细胞相当的生理屏障功能。显示生理屏障功能的微血管在I型胶原水凝胶中形成，其硬度与人脑的硬度相匹配。微血管的扩张反应随跨壁压力的增加呈线性关系，并依赖于基质硬度。这些结果共同提高了组织工程化血脑屏障模型的能力。
Three-dimensional (3D) tissue-engineered models of the blood-brain barrier (BBB) recapitulate in vivo shear stress, cylindrical geometry, and cell-ECM interactions. Here we address four issues associated with BBB models: cell source, barrier function, cryopreservation, and matrix stiffness. We reproduce a directed differentiation of brain microvascular endothelial cells (dhBMECs) from two fluorescently labeled human induced pluripotent stem cell lines (hiPSCs) and demonstrate physiological permeability of Lucifer yellow over six days. Microvessels formed from cryopreserved dhBMECs show expression of BBB markers and maintain physiological barrier function comparable to non-cryopreserved cells. Microvessels displaying physiological barrier function are formed in collagen I hydrogels with stiffness matching that of human brain. The dilation response of microvessels was linear with increasing transmural pressure and was dependent on matrix stiffness. Together these results advance capabilities for tissue-engineered BBB models.