Mechanotransduction is widely used to guide cell fate in hydrogels. Traditionally, hydrogels contain adhesive ligands that dynamically bond with cells to stimulate biochemical signaling axes such as YAP‐TAZ. However, the molecular toolbox to achieve mechanotransduction remains virtually limited to non‐covalent bonds, which limits the ability to program engineered living matter. Here, it is demonstrated that on‐cell chemistry can be leveraged to covalently tether biomaterials directly onto cells, that reveal mechanotransduction via intracellular biophysical programming. Specifically, droplet microfluidics is used to produce single‐cell microgels in which individual stem cells are covalently tethered to either soft or stiff hydrogels via on‐cell oxidative phenolic coupling. Investigation of mechanotransduction effects at single‐cell resolution reveals altered intracellular molecular crowding, calcium signaling, and chromatin organization by regulating cytoplasmic and nuclear volume in a stiffness‐dependent yet YAP/TAZ‐independent manner. Notably, the addition of conventional dynamic adhesive ligands such as RGDs decreases the chondrogenic commitment of stem cells indicating that covalent cell‐material tethering is both efficient and sufficient for programming cell fate. Hence, encoding biomaterials onto cells using covalent on‐cell chemistry to attain mechanotransduction expands the ability to guide cellular behavior, which can accelerate the development of in vitro drug‐screening models, biofabrication of lab‐grown meat, and engineered tissues.

中文翻译：

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通过氧化酚类偶联物结合生物材料的共价细胞内偶联通过细胞内生物物理编程调节干细胞命运


机械转导广泛用于指导水凝胶中的细胞命运。传统上，水凝胶包含粘附配体，这些配体与细胞动态结合以刺激生化信号轴，例如 YAP-TAZ。然而，实现机械转导的分子工具箱实际上仍然局限于非共价键，这限制了对工程生命物质进行编程的能力。在这里，证明可以利用细胞内化学将生物材料直接共价系留到细胞上，从而通过细胞内生物物理编程揭示机械转导。具体来说，液滴微流控用于生产单细胞微凝胶，其中单个干细胞通过细胞上氧化酚类偶联共价拴系在软水凝胶或硬水凝胶上。在单细胞分辨率下对机械转导效应的研究揭示了通过以刚度依赖性但 YAP/TAZ 非依赖性的方式调节细胞质和细胞核体积，改变了细胞内分子拥挤、钙信号传导和染色质组织。值得注意的是，添加常规动态粘附配体（如 RGD）会降低干细胞的软骨形成定型，这表明共价细胞-材料栓系对于编程细胞命运既有效又足够。因此，使用共价细胞上化学将生物材料编码到细胞上以实现机械转导，扩展了指导细胞行为的能力，这可以加速体外药物筛选模型的开发、实验室培育肉的生物制造和工程组织的开发。