Matrix Composition and Mechanics Direct Proangiogenic Signaling from Mesenchymal Stem Cells
Matrix Composition and Mechanics Direct Proangiogenic Signaling from Mesenchymal Stem Cells
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DOI：
10.1089/ten.tea.2013.0661
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发表时间：
2014-10-01
影响因子：
4.1
通讯作者：
Kilian, Kristopher A.
中科院分区：
文献类型：
--
作者：
Abdeen, Amr A.;Weiss, Jared B.;Kilian, Kristopher A.
从间充质干细胞（MSC）分泌促进血管生成的营养因子是一种有前途的基于细胞的治疗方法。然而，临床疗效已被证明是可变的，可能是由于不明确的细胞递送制剂和细胞分泌的固有复杂性。在这里，我们展示了如何控制MSC周围细胞外基质（ECM）的机械性能和蛋白质组成可以引导促血管生成信号。将粘附于用纤连蛋白、胶原蛋白I或层粘连蛋白功能化的聚丙烯酰胺水凝胶的MSC的条件培养基应用于含有人微血管内皮细胞（HMVEC）的3D基质胶培养物。HMVECs中的微管形成的程度取决于底物的刚性和基质蛋白的组成。在纤连蛋白改性的水凝胶上培养的MSC显示出促血管生成信号传导的刚度依赖性，其中从40 kPa条件培养基观察到对小管形成的最大影响，比市售的生长因子混合物高两倍。定量实时聚合酶链反应揭示了参与血管生成的多种因子的刚性依赖性表达，证实了功能性小管生成测定。用微图案化表面限制细胞扩散减弱了条件培养基的作用;然而，肌动球蛋白收缩性的小分子抑制剂不会显著降低功能结果。这项工作证明了如何控制基质刚度和蛋白质组成可以影响间充质干细胞的分泌概况。解构MSC分泌中涉及的物理和生化线索的模型系统可能有助于设计用于基于细胞的治疗的水凝胶生物材料。
The secretion of trophic factors that promote angiogenesis from mesenchymal stem cells (MSCs) is a promising cell-based therapeutic treatment. However, clinical efficacy has proved variable, likely on account of ill-defined cell delivery formulations and the inherent complexity of cellular secretion. Here we show how controlling the mechanical properties and protein composition of the extracellular matrix (ECM) surrounding MSCs can guide proangiogenic signaling. Conditioned media from MSCs adherent to polyacrylamide hydrogel functionalized with fibronectin, collagen I, or laminin was applied to 3D matrigel cultures containing human microvascular endothelial cells (HMVECs). The degree of tubulogenesis in HMVECs is shown to depend on both the substrate rigidity and matrix protein composition. MSCs cultured on fibronectin-modified hydrogels show a stiffness dependence in proangiogenic signaling with maximum influence on tubulogenesis observed from 40 kPa conditioned media, twofold higher than commercially available cocktails of growth factors. Quantitative real-time-polymerase chain reaction reveals stiffness-dependent expression of multiple factors involved in angiogenesis that corroborate the functional tubulogenesis assay. Restricting cell spreading with micropatterned surfaces attenuates the conditioned media effects; however, small-molecule inhibitors of actomyosin contractility do not significantly reduce the functional outcome. This work demonstrates how controlling matrix rigidity and protein composition can influence the secretory profile of MSCs. Model systems that deconstruct the physical and biochemical cues involved in MSC secretion may assist in the design of hydrogel biomaterials for cell-based therapies.