Functionality of Endothelial Cells and Pericytes From Human Pluripotent Stem Cells Demonstrated in Cultured Vascular Plexus and Zebrafish Xenografts
Functionality of Endothelial Cells and Pericytes From Human Pluripotent Stem Cells Demonstrated in Cultured Vascular Plexus and Zebrafish Xenografts
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DOI：
10.1161/atvbaha.113.302598
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发表时间：
2014-01-01
影响因子：
8.7
通讯作者：
Mummery, Christine L.
中科院分区：
文献类型：
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作者：
Orlova, Valeria V.;Drabsch, Yvette;Mummery, Christine L.
目的：血管内皮细胞(ECs)、周细胞和血管平滑肌细胞(VSMCs)是血管发育所必需的，其功能障碍可导致多种心血管疾病。然而，用于研究的原代血管细胞很难获得。从体细胞来源的人诱导多能干细胞(HiPSCs)是一种可再生的ECs和vSMC来源；然而，由于分化效率低且不一致以及缺乏表型生物测定，它们作为疾病模型的使用一直受到限制。方法和结果-在这里，我们定义了从不同组织来源的hiPSCs同时高效地获得ECs和周细胞的条件。该方案对所有品系和人类胚胎干细胞(HESCs)都同样有效。内皮细胞可连续传代，表型难以区分，表现出动脉样胚胎内皮细胞的特征。此外，当HiPSC来源的内皮细胞与HiPSC来源的周细胞共同培养时，形成了真正的血管丛。共培养体系概括了(1)血管发育的主要步骤，包括EC增殖和原发神经丛重塑；(2)EC介导的成熟和周细胞获得收缩的vSMC表型。此外，HiPSC来源的内皮细胞作为异种移植整合到斑马鱼发育中的血管系统中。这与更广泛使用的人脐静脉内皮细胞相比，后者只能形成不稳定的血管，完全无法整合到斑马鱼血管中。结论-我们证明了HiPSC的血管衍生物，如内皮细胞和周细胞，具有完整的功能，可以用于体外研究缺陷内皮细胞与周细胞的相互作用，用于疾病建模和肿瘤血管生成的研究。
Objective-Endothelial cells (ECs), pericytes, and vascular smooth muscle cells (vSMCs) are essential for vascular development, and their dysfunction causes multiple cardiovascular diseases. Primary vascular cells for research are, however, difficult to obtain. Human-induced pluripotent stem cells (hiPSCs) derived from somatic tissue are a renewable source of ECs and vSMCs; however, their use as disease models has been limited by low and inconsistent efficiencies of differentiation and the lack of phenotypic bioassays.Approach and Results-Here, we developed defined conditions for simultaneous derivation of ECs and pericytes with high efficiency from hiPSCs of different tissue origin. The protocol was equally efficient for all lines and human embryonic stem cells (hESCs). The ECs could undergo sequential passage and were phenotypically indistinguishable, exhibiting features of arterial-like embryonic ECs. Moreover, hiPSC-derived ECs formed an authentic vascular plexus when cocultured with hiPSC-derived pericytes. The coculture system recapitulated (1) major steps of vascular development including EC proliferation and primary plexus remodeling, and (2) EC-mediated maturation and acquisition of contractile vSMC phenotype by pericytes. In addition, hiPSC-derived ECs integrated into developing vasculature as xenografts in zebrafish. This contrasts with more widely used ECs from human umbilical vein, which form only unstable vasculature and were completely unable to integrate into zebrafish blood vessels.Conclusions-We demonstrate that vascular derivatives of hiPSC, such as ECs and pericytes, are fully functional and can be used to study defective endothelia-pericyte interactions in vitro for disease modeling and studies on tumor angiogenesis.