The dynamic migratory behavior of human mesenchymal stem cells (hMSCs) has a significant impact on the epigenetic profiles that determine fate choice and lineage commitment during differentiation. Here we report a novel approach to enhance repeated migration-driven epigenetic memory which induces cardiomyogenic differentiation on a dendrimer surface with fifth generation (G5). Cells exhibited the formation of cell aggregates on the G5 surface through active migration with morphological changes, and these aggregates showed strong expression of the cardiac-specific marker cardiac troponin T (cTnT) at 10 days. When cell aggregates were passaged onto a fresh G5 surface over three passages of 40 days, the expression levels of the multiple cardiac-specific markers including GATA4, NKX2.5, MYH7, and TNNT2 were higher compared to those passaged as single cells. To investigate whether cardiomyogenic differentiation of hMSCs was enhanced by repeated aggregate migration-driven epigenetic memory, cells on the G5 surface were reseeded onto a fresh G5 surface during three passages using aggregate-based and single cell-based passage methods. Analyses of global changes in H3 histone modifications exhibited pattern of increased H3K9ac and H3K27me3, and decreased H3K9me3 in aggregate-based passage cultures during three passages. However, the pattern of their histone modification on the PS surface was repeated after the initialization and reformation during three passages in single cell-based passage cultures. Thus, repetitive aggregate migratory behavior during aggregate-based passage led to a greater degree of histone modification, as well as gene expression changes suggestive of cardiomyogenic differentiation.

人类间充质干细胞 (hMSCs) 的动态迁移行为对决定分化过程中命运选择和谱系承诺的表观遗传谱有重大影响。在这里，我们报告了一种增强重复迁移驱动的表观遗传记忆的新方法，该方法在第五代 (G5) 的树枝状聚合物表面诱导心肌分化。细胞通过形态变化的主动迁移在 G5 表面形成细胞聚集体，这些聚集体在 10 天时表现出心脏特异性标志物心肌肌钙蛋白 T (cTnT) 的强烈表达。当细胞聚集体在 3 次 40 天内传代到新鲜的 G5 表面时，包括 GATA4、NKX2.5、MYH7、与作为单细胞传代的那些相比，TNNT2 和 TNNT2 更高。为了研究重复的聚集迁移驱动的表观遗传记忆是否增强了 hMSC 的心肌细胞分化，使用基于聚集和基于单细胞的传代方法在三个传代过程中将 G5 表面上的细胞重新接种到新鲜的 G5 表面上。H3 组蛋白修饰的整体变化分析表明，在三个传代期间，基于聚合的传代培养物中 H3K9ac 和 H3K27me3 增加的模式和 H3K9me3 减少的模式。然而，在基于单细胞的传代培养的三个传代中，在初始化和重组后，PS 表面上的组蛋白修饰模式被重复。因此，在基于聚合的传代过程中重复的聚合迁移行为导致了更大程度的组蛋白修饰，