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：
10.1002/mabi.202300110
复制
发表时间：
2024
影响因子：
4.6
通讯作者：
Caliari, Steven R.
中科院分区：
文献类型：
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作者：
Sumey, Jenna L.;Harrell, Abigail M.;Johnston, Peyton C.;Caliari, Steven R.
人们对开发水凝胶作为可调的体外细胞培养平台以研究细胞以受控方式对机械刺激的反应有极大的兴趣。然而，很少有人知道常见的细胞培养技术，如组织培养塑料上的连续扩增，如何影响随后的细胞在水凝胶上培养时的行为。在这项工作中，利用甲基丙烯酸化透明质酸水凝胶平台来研究基质细胞的机械转导。水凝胶首先通过硫醇-迈克尔加成形成，以模拟正常软组织（例如，肺）僵硬度（E ≥ 1 kPa）。通过未消耗的甲基丙烯酸酯的自由基光聚合进行二次交联，可以匹配早期（E ≤ 6 kPa）和晚期纤维化组织（E ≤ 50 kPa）。早期传代（P1）的人骨髓间充质基质细胞（hMSC）显示随着水凝胶硬度的增加，扩散、心肌细胞因子相关转录因子-A（MRTF-A）核定位和粘着斑大小增加。然而，与早期传代hMSC相比，晚期传代（P5）hMSC显示出对基质力学的敏感性降低，具有较低的MRTF-A核转位和在较硬的水凝胶上较小的局灶性粘连。在永生化人肺成纤维细胞系中观察到类似的趋势。总的来说，这项工作突出了标准细胞培养实践的影响，研究细胞响应机械信号使用体外水凝胶模型。
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.