Zhang Fan, Zheng Lu, Wu Yi, Ding Qihan, Lü Shouqin, Lü Dongyuan, Long Mian
Center for Biomechanics and Bioengineering, Beijing Key Laboratory of Engineered Construction and Mechanobiology and Key Laboratory of Microgravity, Institute of Mechanics, Chinese Academy of Sciences, Beijing, 100190, China.
School of Engineering Science, University of Chinese Academy of Sciences, Beijing, 100049, China.
Cell Mol Life Sci. 2025 Jun 23;82(1):245. doi: 10.1007/s00018-025-05776-9.
Intercellular adhesion is crucial in regulating stemness maintenance and differentiation initiation of embryonic stem cells (ESCs), which is also cooperated with extracellular mechanical microenvironment. Here an in vitro model was used to elucidate the effects of E-cadherin complexes on definitive endoderm (DE)-directed differentiation of hESCs (H1 cells), when the cells were seeded on polyacrylamide hydrogels with varied stiffnesses. Results indicated that stiff substrate increased the proportion of H1 cells differentiating into DE cells and intercellular E-cadherin expression was reduced with progressive stages at same stiffness, presenting a negative correlation of E-cadherin expression with differentiating progress or substrate stiffness. Blocking E-cadherin enhanced the productivity of differentiated cells and promoted the disassembly of intercellular adhesions by translocating YAP into nuclei, which was positively correlated with GATA6 and CXCR4 expressions in a stiffness-dependent manner. This work provided an insight into understanding the roles of E-cadherin-related intercellular adhesion and substrate stiffness in DE-directed differentiation of hESCs.
细胞间黏附在调节胚胎干细胞（ESC）干性维持和分化起始过程中至关重要，且其与细胞外机械微环境协同作用。在此，当细胞接种在具有不同硬度的聚丙烯酰胺水凝胶上时，利用体外模型阐明E-钙黏蛋白复合物对人胚胎干细胞（H1细胞）向定形内胚层（DE）分化的影响。结果表明，硬底物增加了H1细胞分化为DE细胞的比例，且在相同硬度条件下，随着分化进程的推进，细胞间E-钙黏蛋白表达降低，呈现出E-钙黏蛋白表达与分化进程或底物硬度的负相关。阻断E-钙黏蛋白可提高分化细胞的产量，并通过将YAP转运至细胞核促进细胞间黏附的解离，这与GATA6和CXCR4表达呈刚度依赖性正相关。这项工作为理解E-钙黏蛋白相关的细胞间黏附和底物硬度在人胚胎干细胞向DE分化中的作用提供了见解。
