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首页> 外文期刊>Scientific reports. >An immortalised mesenchymal stem cell line maintains mechano-responsive behaviour and can be used as a reporter of substrate stiffness
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An immortalised mesenchymal stem cell line maintains mechano-responsive behaviour and can be used as a reporter of substrate stiffness

机译：永生化的间充质干细胞系保持机械响应性行为，可用作基材刚度的报告

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The mechanical environment can influence cell behaviour, including changes to transcriptional and proteomic regulation, morphology and, in the case of stem cells, commitment to lineage. However, current tools for characterizing substrates’ mechanical properties, such as atomic force microscopy (AFM), often do not fully recapitulate the length and time scales over which cells ‘feel’ substrates. Here, we show that an immortalised, clonal line of human mesenchymal stem cells (MSCs) maintains the responsiveness to substrate mechanics observed in primary cells, and can be used as a reporter of stiffness. MSCs were cultured on soft and stiff polyacrylamide hydrogels. In both primary and immortalised MSCs, stiffer substrates promoted increased cell spreading, expression of lamin-A/C and translocation of mechano-sensitive proteins YAP1 and MKL1 to the nucleus. Stiffness was also found to regulate transcriptional markers of lineage. A GFP-YAP/RFP-H2B reporter construct was designed and virally delivered to the immortalised MSCs for in situ detection of substrate stiffness. MSCs with stable expression of the reporter showed GFP-YAP to be colocalised with nuclear RFP-H2B on stiff substrates, enabling development of a cellular reporter of substrate stiffness. This will facilitate mechanical characterisation of new materials developed for applications in tissue engineering and regenerative medicine.
机译：机械环境可以影响细胞行为，包括转录和蛋白质组学调节，形态的变化，在干细胞的情况下，对谱系的承诺。然而，用于表征基板的机械性能的电流工具，例如原子力显微镜（AFM），通常不会完全重新承载细胞“感觉”基板的长度和时间尺度。这里，我们表明，一种永生化的人间充质干细胞（MSCs）的克隆线保持对原发性细胞中观察到的基材力学的反应性，并且可以用作刚度的报告。在柔软和硬丙烯酰胺水凝胶上培养MSC。在初级和永生化的MSC中，致纤维底物促进了增加的细胞扩散，Lamin-A / C的表达以及机械敏感蛋白Yap1和MkL1的易位到细胞核。还发现刚度调节谱系的转录标志物。设计并在底物刚度的原位检测中设计并公用地递送到永生的MSCs的GFP-YAP / RFP-H2B报告构建体。具有稳定表达报告的MSC显示出GFP-yap在硬质基板上用核RFP-H2B结合，从而能够开发基质刚度的细胞报告。这将促进为组织工程和再生医学应用开发的新材料的机械表征。

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