Serial passaging affects stromal cell mechanosensitivity on hyaluronic acid hydrogels.
Serial passaging affects stromal cell mechanosensitivity on hyaluronic acid hydrogels.
复制标题
连续传代影响基质细胞对透明质酸水凝胶的机械敏感性。
DOI：
10.1101/2023.03.16.532853
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Caliari,StevenR
中科院分区：
文献类型：
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作者：
Sumey,JennaL;Harrell,AbigailM;Johnston,PeytonC;Caliari,StevenR
开发水凝胶作为可调的体外细胞培养平台，以受控的方式研究细胞对机械线索的反应是非常有兴趣的。然而，对于常见的细胞培养技术，如组织培养塑料上的连续扩增，在水凝胶上培养时如何影响细胞的后续行为，人们知之甚少。在这项工作中，甲基丙烯酸透明质酸水凝胶平台被用来研究基质细胞的机械转导。水凝胶首先通过硫醇- Michael添加到模型正常软组织（如肺）刚度（E≈1 kPa）中形成。未消耗的甲基丙烯酸酯通过自由基光聚合进行二次交联，可实现早期（E≈6 kPa）和晚期纤维化组织（E≈50 kPa）的匹配。随着水凝胶硬度的增加，早期传代（P1）人骨髓间充质间质细胞（hMSCs）的扩散、心肌素相关转录因子- A （MRTF - A）核定位和局灶黏附大小增加。然而，与早期传代hMSCs相比，晚期传代（P5） hMSCs对底物力学的敏感性降低，MRTF‐A核易位降低，在更硬的水凝胶上的黏附更小。在永生化的人肺成纤维细胞系中也观察到类似的趋势。总的来说，这项工作强调了使用体外水凝胶模型研究细胞对机械信号反应的标准细胞培养实践的意义。
There is a tremendous interest in developing hydrogels as tunable in vitro cell culture platforms to study cell response to mechanical cues in a controlled manner. However, little is known about how common cell culture techniques, such as serial expansion on tissue culture plastic, affect subsequent cell behavior when cultured on hydrogels. In this work, a methacrylated hyaluronic acid hydrogel platform is leveraged to study stromal cell mechanotransduction. Hydrogels are first formed through thiol‐Michael addition to model normal soft tissue (e.g., lung) stiffness (E≈ 1 kPa). Secondary cross‐linking via radical photopolymerization of unconsumed methacrylates allows matching of early‐ (E≈ 6 kPa) and late‐stage fibrotic tissue (E≈ 50 kPa). Early passage (P1) human bone marrow mesenchymal stromal cells (hMSCs) display increased spreading, myocardin‐related transcription factor‐A (MRTF‐A) nuclear localization, and focal adhesion size with increasing hydrogel stiffness. However, late passage (P5) hMSCs show reduced sensitivity to substrate mechanics with lower MRTF‐A nuclear translocation and smaller focal adhesions on stiffer hydrogels compared to early passage hMSCs. Similar trends are observed in an immortalized human lung fibroblast line. Overall, this work highlights the implications of standard cell culture practices on investigating cell response to mechanical signals using in vitro hydrogel models.