Peptide modified nanofibrous scaffold promotes human mesenchymal stem cell proliferation and long-term passaging.
Peptide modified nanofibrous scaffold promotes human mesenchymal stem cell proliferation and long-term passaging.
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DOI：
10.1016/j.msec.2017.11.017
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发表时间：
2018-03
期刊：
影响因子：
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通讯作者：
Rezvan Mobasseri;Lingling Tian;M. Soleimani;S. Ramakrishna;H. Naderi-manesh
中科院分区：
文献类型：
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作者：
Rezvan Mobasseri;Lingling Tian;M. Soleimani;S. Ramakrishna;H. Naderi-manesh
人骨髓间充质干细胞（human mesenchymal stem cells，hMSCs）长期培养、传代和增殖导致其干细胞特性（包括自我更新和多能性）的丧失。通过优化MSCs的体外培养环境，维持干细胞状态和更好地控制细胞命运是可能的。我们最近报道了生物活性达特蛋白衍生肽R-肽对hMSC粘附、形态和增殖的显著影响，与其他生物活性分子包括RGD肽、纤连蛋白和胶原相比，R-肽增强了MSC的早期粘附和增殖。在这项研究中，R-肽用于评估长期传代后的MSC的干性特性。以R肽修饰的聚己内酯（PCL）纳米纤维和未修饰的PCL纳米纤维为支架材料，研究了R肽修饰的PCL纳米纤维和PCL纳米纤维对细胞行为的影响。结果显示，细胞接种后30 min，R-肽修饰的支架上早期形成粘着斑（FA）复合物。由于R-肽的存在，细胞增殖速率显著增加，并且使用流式细胞术和免疫细胞化学染色的MSC标记物分析证明了R-肽即使在长期传代培养后也能够维持间充质干细胞特性（高增殖、多能标记物表达和分化能力）。因此，我们的结果得出结论，生物活性R-肽与纳米纤维支架的组合可以以最佳方式模拟包含微/纳米结构和生物化学分子的天然ECM。所设计的支架可以通过FA的形成和细胞骨架的组织将细胞外基质（ECM）连接到细胞核，引起MSC的快速和牢固的附着，并允许整合素介导的信号传导开始。
Long-term culture, passage and proliferation of human mesenchymal stem cells (hMSCs) cause loss of their stemness properties including self-renewal and multipotency. By optimizing the MSCs environment in vitro, maintaining the stemness state and better controlling the cell fate might be possible. We have recently reported the significant effects of bioactive Tat protein-derived peptide named R-peptide on hMSC adhesion, morphology and proliferation, which has demonstrated R-peptide enhanced MSC early adhesion and proliferation in comparison to other bioactive molecules including RGD peptide, fibronectin and collagen. In this study, R-peptide was used to evaluate stemness properties of MSCs after long-term passaging. R-peptide conjugated poly caprolactone (PCL) nanofibrous scaffold and unmodified nanofibrous scaffold were used to study the impact of R-peptide modified PCL nanofibers and PCL nanofibers on cell behavior. The results showed early formation of focal adhesion (FA) complex on R-peptide modified scaffolds at 30 min after cell seeding. The rate of cell proliferation was significantly increased due to presence of R-peptide, and the MSCs marker analyses using flow cytometry and immunocytochemistry staining proved the ability of R-peptide to maintain mesenchymal stem cell properties (high proliferation, expression of multipotent markers and differentiation capacity) even after long-term passage culturing. Accordingly, our (The) results concluded that bioactive R-peptide in combination with nanofibrous scaffold can mimic the native ECM comprising micro/nano architecture and biochemical molecules in a best way. The designed scaffold can link extracellular matrix (ECM) to nucleus via formation of FA and organization of cytoskeleton, causing fast and strong attachment of MSCs and allowing integrin-mediated signaling to start.