From the Department of Anatomy and Embryology (V.V.O., C.F., S.P.-R., F.E.v.d.H., S.M., C.L.M.) and Department of Molecular Cell Biology (V.V.O., Y.D., P.t.D.), Cancer Genomics Centre and Centre for Biomedical Genetics, Leiden University Medical Center, Leiden, The Netherlands.
Arterioscler Thromb Vasc Biol. 2014 Jan;34(1):177-86. doi: 10.1161/ATVBAHA.113.302598. Epub 2013 Oct 24.
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.
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.
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.
内皮细胞（ECs）、周细胞和血管平滑肌细胞（vSMCs）对于血管发育至关重要，其功能障碍会导致多种心血管疾病。然而，用于研究的主要血管细胞难以获得。源自体组织的人诱导多能干细胞（hiPSCs）是 ECs 和 vSMCs 的可再生来源；然而，由于分化效率低且不一致，以及缺乏表型生物测定，其作为疾病模型的应用受到限制。
在这里，我们开发了从不同组织来源的 hiPSCs 高效同时衍生 ECs 和周细胞的定义条件。该方案对所有细胞系和人类胚胎干细胞（hESCs）同样有效。ECs 可以进行连续传代，表型上无明显差异，表现出类似于动脉样胚胎 ECs 的特征。此外，hiPSC 衍生的 ECs 在与 hiPSC 衍生的周细胞共培养时形成了真实的血管丛。共培养系统再现了（1）血管发育的主要步骤，包括 EC 增殖和初级丛状结构重塑，以及（2）EC 介导的周细胞成熟和获得收缩性 vSMC 表型。此外，hiPSC 衍生的 ECs 作为异种移植物整合到斑马鱼发育中的血管中。这与更广泛使用的源自人脐静脉的 ECs 形成鲜明对比，后者仅形成不稳定的血管，并且完全无法整合到斑马鱼血管中。
我们证明了 hiPSC 的血管衍生物，如 ECs 和周细胞，具有完全的功能，可以用于体外研究内皮细胞-周细胞相互作用的缺陷，用于疾病建模和肿瘤血管生成研究。
