Center for Biomechanics and Bioengineering, Key Laboratory of Microgravity (National Microgravity Laboratory) and Beijing Key Laboratory of Engineered Construction and Mechanobiology, Institute of Mechanics, Chinese Academy of Sciences, Beijing 100190, China.
CAS Key Laboratory of Molecular Imaging, Institute of Automation, Chinese Academy of Sciences, Beijing 100190, China.
Biomater Sci. 2021 May 18;9(10):3776-3790. doi: 10.1039/d1bm00174d.
Mechanical or physical cues are associated with the growth and differentiation of embryonic stem cells (ESCs). While the substrate stiffness or topography independently affects the differentiation of ESCs, their cooperative regulation on lineage-specific differentiation remains largely unknown. Here, four topographical configurations on stiff or soft polyacrylamide hydrogel were combined to direct hepatic differentiation of human H1 cells via a four-stage protocol, and the coupled impacts of stiffness and topography were quantified at distinct stages. Data indicated that the substrate stiffness is dominant in stemness maintenance on stiff gel and hepatic differentiation on soft gel while substrate topography assists the differentiation of hepatocyte-like cells in positive correlation with the circularity of H1 clones initially formed on the substrate. The differentiated cells exhibited liver-specific functions such as maintaining the capacities of CYP450 metabolism, glycogen synthesis, ICG engulfment, and repairing liver injury in CCl4-treated mice. These results implied that the coupling of substrate stiffness and topography, combined with the biochemical signals, is favorable to improve the efficiency and functionality of hepatic differentiation of human ESCs.
机械或物理线索与胚胎干细胞（ESCs）的生长和分化有关。虽然底物的硬度或形貌独立地影响 ESCs 的分化，但它们对线粒体特异性分化的协同调节在很大程度上仍然未知。在这里，通过四阶段方案，将硬或软聚丙烯酰胺水凝胶上的四个形貌组合起来，以指导人 H1 细胞的肝向分化，并在不同阶段量化了刚度和形貌的耦合影响。数据表明，在硬凝胶上维持干细胞特性和软凝胶上进行肝分化时，基质硬度起主要作用，而基质形貌通过与最初在基质上形成的 H1 克隆的圆度呈正相关，有助于肝样细胞的分化。分化细胞表现出肝脏特异性功能，如保持 CYP450 代谢、糖原合成、ICG 吞噬和修复 CCl4 处理的小鼠肝损伤的能力。这些结果表明，基质硬度和形貌的结合以及生化信号的结合有利于提高人 ESC 肝向分化的效率和功能。
