Tissue-specific extracellular matrix promotes myogenic differentiation of human muscle progenitor cells on gelatin and heparin conjugated alginate hydrogels.
Tissue-specific extracellular matrix promotes myogenic differentiation of human muscle progenitor cells on gelatin and heparin conjugated alginate hydrogels.
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DOI：
10.1016/j.actbio.2017.08.022
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发表时间：
2017-10-15
影响因子：
9.7
通讯作者：
Zhang Y
中科院分区：
文献类型：
--
作者：
Yi H;Forsythe S;He Y;Liu Q;Xiong G;Wei S;Li G;Atala A;Skardal A;Zhang Y
骨骼肌祖细胞（SMPCs）的成肌分化、细胞融合和肌管形成在骨骼肌发育和修复过程中起着关键作用。然而，在从活组织分离并转移到培养皿后，SMPC由于细胞外基质（ECM）的缺乏而逐渐丧失其功能并停止繁殖。尽管在细胞培养物中使用ECM组分以维持和繁殖某些组织类型的实验结果令人鼓舞，但这种方法对SMPC培养的益处是有限的，因为生物活性分子和蛋白质在培养期间立即释放并降解。在这项研究中，我们开发了一种新的方法，以提高人骨骼肌祖细胞（hSMPCs）的增殖和分化的骨骼肌ECM结合与明胶和肝素（Alg-G-H）作为基板的改性藻酸盐水凝胶在体外。与单独的基质（即明胶、Matrigel®或单独的ECM）相比，该Alg-G-H底物与骨骼肌ECM一起显著增强hSMPC的细胞扩增、分化和成熟。在蛋白质印迹和免疫细胞化学分析中，Alg-G-H-ECM主要增强骨骼肌发生标志物（MyoD、Myf 5、肌细胞生成素、结蛋白和肌球蛋白）的表达和hSMPC中的肌管形成。该研究表明，将Alg-G-H底物与骨骼肌ECM组合通过释放信号分子和生长因子来调节hSMPCs的细胞增殖、分化和成熟的稳态。该技术可能是体外骨骼肌细胞分化和成熟的一种具有成本效益的工具，在组织再生和药物开发中具有潜在的应用前景。
Myogenic differentiation, cell fusion, and myotube formation of skeletal muscle progenitor cells (SMPCs) have key roles during skeletal muscle development and repair. However, after isolation from living tissue and transition to culture dishes, SMPCs gradually lose their function and stop propagating due to the absence of extracellular matrix (ECM). Despite encouraging results of experiments using ECM components in cell culture for maintenance and propagation of some tissue types, the benefits of this approach on SMPC culture are limited, because the bioactive molecules and proteins instantly release and are degraded during culture. In this study, we developed a novel approach to enhance the proliferation and differentiation of human skeletal muscle progenitor cells (hSMPCs) in vitro with skeletal muscle ECM in combination with a modified alginate hydrogel conjugated with gelatin and heparin (Alg-G-H) as a substrate. This Alg-G-H substrate, together with skeletal muscle ECM, significantly enhanced cell expansion, differentiation, and maturation of hSMPCs compared with individual substrata (i.e. gelatin, Matrigel®, or ECM alone). In Western-blot and immunocytochemical analyses, the Alg-G-H-ECM predominantly enhanced expression of skeletal myogenesis markers (MyoD, Myf5, Myogenin, Desmin and Myosin) and myotube formation in hSMPCs. This study demonstrated that combining Alg-G-H substrates with skeletal muscle ECM modulated homeostasis of cell proliferation, differentiation, and maturation of hSMPCs by releasing signaling molecules and growth factors. This technique could be a cost-effective tool for in vitro skeletal muscle cell differentiation and maturation, with potential applications in tissue regeneration and drug development.
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DOI：
10.1002/adma.201600240
发表时间：
2016-12
期刊：
Advanced materials (Deerfield Beach, Fla.)
影响因子：
--
作者：
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发表时间：
2015-01
期刊：
Journal of materials science. Materials in medicine
影响因子：
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作者：
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影响因子：
3
作者：
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通讯作者：
Ribchester, R. R.
影响因子：
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