Functional vascular smooth muscle cells derived from human induced pluripotent stem cells via mesenchymal stem cell intermediates
Functional vascular smooth muscle cells derived from human induced pluripotent stem cells via mesenchymal stem cell intermediates
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DOI：
10.1093/cvr/cvs253
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发表时间：
2012-12-01
影响因子：
10.8
通讯作者：
Andreadis, Stelios T.
中科院分区：
文献类型：
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作者：
Bajpai, Vivek K.;Mistriotis, Panagiotis;Andreadis, Stelios T.
平滑肌细胞（SMC）在血管稳态和疾病中发挥着重要作用。尽管成体间充质干细胞 (MSC) 已被用作收缩性 SMC 的来源，但它们的增殖潜力有限且培养衰老，特别是来自老年供体时。相比之下，人类诱导多能干细胞 (hiPSC) 可以为基于自体细胞的疗法和创建血管疾病模型提供无限的功能性 SMC 来源。我们的目标是开发一种有效的策略，从 hiPSC 中衍生出功能性、收缩性 SMC。我们开发了一种稳健、分阶段、无饲养层的策略，通过多能 MSC 的中间阶段将 hiPSC 分化为功能性 SMC，可以诱导分化为脂肪、骨骼、软骨和肌肉。在此阶段，与亲本毛囊间充质干细胞相比，细胞高度增殖，表现出更高的克隆形成潜力，并且衰老减少。此外，当暴露于分化培养基时，肌源蛋白，例如平滑肌肌动蛋白、钙调蛋白和肌球蛋白重链显着上调，并表现出强大的纤维组织，表明收缩表型的发展。事实上，由这些细胞制备的组织构建体对受体和非受体介导的激动剂表现出高水平的收缩性。我们开发了一种有效的分阶段策略，使 hiPSC 通过克隆和多能 MSC 的中间群体分化为收缩性 SMC。 MSC 和 SMC 衍生的高产量表明我们的策略可能有助于获得再生医学或研究血管疾病病理生理学所需的大量细胞。
Smooth muscle cells (SMC) play an important role in vascular homeostasis and disease. Although adult mesenchymal stem cells (MSC) have been used as a source of contractile SMC, they suffer from limited proliferation potential and culture senescence, particularly when originating from older donors. By comparison, human induced pluripotent stem cells (hiPSC) can provide an unlimited source of functional SMC for autologous cell-based therapies and for creating models of vascular disease. Our goal was to develop an efficient strategy to derive functional, contractile SMC from hiPSC.We developed a robust, stage-wise, feeder-free strategy for hiPSC differentiation into functional SMC through an intermediate stage of multipotent MSC, which could be coaxed to differentiate into fat, bone, cartilage, and muscle. At this stage, the cells were highly proliferative and displayed higher clonogenic potential and reduced senescence when compared with parental hair follicle mesenchymal stem cells. In addition, when exposed to differentiation medium, the myogenic proteins such as -smooth muscle actin, calponin, and myosin heavy chain were significantly upregulated and displayed robust fibrillar organization, suggesting the development of a contractile phenotype. Indeed, tissue constructs prepared from these cells exhibited high levels of contractility in response to receptor- and non-receptor-mediated agonists.We developed an efficient stage-wise strategy that enabled hiPSC differentiation into contractile SMC through an intermediate population of clonogenic and multipotent MSC. The high yield of MSC and SMC derivation suggests that our strategy may facilitate an acquisition of the large numbers of cells required for regenerative medicine or for studying vascular disease pathophysiology.