Quantifying the Vasculogenic Potential of Induced Pluripotent Stem Cell-Derived Endothelial Progenitors in Collagen Hydrogels
Quantifying the Vasculogenic Potential of Induced Pluripotent Stem Cell-Derived Endothelial Progenitors in Collagen Hydrogels
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DOI：
10.1089/ten.tea.2018.0274
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发表时间：
2019-05-02
影响因子：
4.1
通讯作者：
Zoldan, Janet
中科院分区：
文献类型：
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作者：
Crosby, Cody O.;Valliappan, Deepti;Zoldan, Janet
诱导多能干细胞衍生的内皮祖细胞（iPSC-EPs）已成为一种有希望的候选细胞来源，用于患者特异性缺血治疗。在这些细胞可以在临床环境中适当部署之前，必须研究它们在细胞外基质(ECM)-模拟，三维（3D）微环境中组装成功能血管网络。为了阐明iPSC-EPs与ECM的相互作用，我们研究了结构蛋白密度的体外调节、血管生成生长因子的存在和相对蛋白水解活性如何影响这些祖细胞的血管生成潜能，即它们自组装成血管样网络的能力。我们发现，添加ROCK途径抑制剂和外源性血管内皮生长因子（VEGF）对于诱导胶原水凝胶中iPSC-EP血管生成至关重要。在这些条件下，含有ve -钙粘蛋白表达管腔的三维血管样网络在培养一周内形成。为了量化这种三维血管样网络，我们开发了一个计算管道来分析网络长度、连通性和平均管腔直径。增加水凝胶中胶原蛋白的浓度会减少网状结构的形成，并促进不相连的大直径管腔的形成。这种现象部分与细胞的蛋白水解能力和水凝胶的性质有关，特别是水凝胶的可变形性和孔径。总之，我们证明iPSC-EPs的血管生成潜能受到细胞-基质相互作用和胶原水凝胶的基质特性的调节。
Induced pluripotent stem cell-derived endothelial progenitors (iPSC-EPs) have emerged as a promising candidate cell source for patient-specific ischemic therapies. Before these cells can be appropriately deployed in a clinical setting, it is imperative to study their assembly into functional vascular networks in extracellular matrix (ECM)-mimicking, three-dimensional (3D) microenvironments. To elucidate the interactions of iPSC-EPs with the ECM, we examined how in vitro modulation of structural protein density, the presence of angiogenic growth factors, and relative proteolytic activity affected the vasculogenic potential of these progenitors, that is, their ability to self-assemble into vessel-like networks. We found that the addition of a ROCK pathway inhibitor and exogenous vascular endothelial growth factor (VEGF) are imperative for inducing robust iPSC-EP vasculogenesis in collagen hydrogels. Under these conditions, 3D vascular-like networks containing VE-cadherin-expressing lumens formed within a week of culture. To quantify this 3D vessel-like network, we developed a computational pipeline to analyze network length, connectivity, and average lumen diameter. Increasing the concentration of collagen in the hydrogels abrogated network formation and encouraged the formation of disconnected, large-diameter lumens. This phenomenon was in part related to the cells' proteolytic capacity and the hydrogels' properties, specifically hydrogel deformability and pore size. In conclusion, we demonstrate that the vasculogenic potential of iPSC-EPs is regulated by cell-matrix interactions and the matrix properties of collagen hydrogels.