Preparation of collagen-coated gels that maximize in vitro myogenesis of stem cells by matching the lateral elasticity of in vivo muscle.
Preparation of collagen-coated gels that maximize in vitro myogenesis of stem cells by matching the lateral elasticity of in vivo muscle.
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DOI：
10.1007/978-1-60761-063-2_13
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Discher, Dennis E
中科院分区：
文献类型：
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作者：
Chaudhuri, Tathagata;Rehfeldt, Florian;Sweeney, H Lee;Discher, Dennis E
细胞微环境的物理性质-包括周围组织的弹性-似乎对细胞形态，细胞骨架和基因表达产生重大影响。我们以前已经表明，只有当细胞生长在与骨骼肌被动顺应性密切匹配的顺应性凝胶上时，定向肌细胞才会形成骨骼肌肌球蛋白II的肌节条纹。我们最近用相同类型的弹性凝胶显示，即使在没有定制的可溶性因子的情况下，间充质干细胞（MSC）也能最大限度地表达肌源性基因。在这里，我们提供了详细的方法，不仅为我们如何制造和纳米力学表征肌肉样弹性的水凝胶，而且我们如何培养间充质干细胞和表征其肌原性诱导全人类基因组转录分析。
The physical nature of a cell’s microenvironment – including the elasticity of the surrounding tissue – appears to exert a significant influence on cell morphology, cytoskeleton, and gene expression. We have previously shown that committed muscle cells will develop sarcomeric striations of skeletal muscle myosin II only when the cells are grown on a compliant gel that closely matches the passive compliance of skeletal muscle. We have more recently shown with the same types of elastic gels that mesenchymal stem cells (MSCs) maximally express myogenic genes, even in the absence of tailored soluble factors. Here, we provide detailed methods not only for how we make and nanomechanically characterize hydrogels of muscle-like elasticity, but also how we culture MSCs and characterize their myogenic induction by whole human genome transcript analysis.