Cardiomyogenic induction of human mesenchymal stem cells by altered Rho family GTPase expression on dendrimer-immobilized surface with d-glucose display

The commitment of stem cells to different lineages is regulated by many cues in the intercellular signals from the microenvironment system. In the present study, we found that alterations in Rho family GTPase activities derived from cytoskeletal formation can lead to guidance of cardiomyogenic differentiation of human mesenchymal stem cells (hMSCs) during in vitro culture. To regulate the cytoskeletal formation of hMSCs, we employed a dendrimer-immobilized substrate that displayed d-glucose. With an increase in the dendrimer generation number, the cells exhibited active migration, accompanied by cell morphological changes of stretching and contracting. Fluorescence microscopy for F-actin, vinculin and glucose transporter1 (GLUT1) clarified the localization of integrin-mediated and GLUT-mediated anchoring, introducing the idea that the morphological changes of the cells were responsive to variations in the generation number of the dendrimer with d-glucose display. On the 5th-generation dendrimer surface, in particular, the cells exhibited RhoA down-regulation and Rac1 up-regulation during the culture, associated with alterations in the cellular morphology and migratory behaviors. It was found that cell aggregation was promoted on this surface, supporting the notion that an increase in N-cadherin-mediated cell–cell contacts and Wnt signaling regulate hMSC differentiation into cardiomyocyte-like cells.