Remodeling Cell Adhesion Releases Cardiac Potential of Human Pluripotent Stem Cells with Continuous Proliferation and Accelerated Maturation.

Human pluripotent stem cells (hPSCs) can generate specific cell types for therapeutic applications. Since cell therapy often requires billions of cells for transplantation, it is essential to maximize differentiation efficiency to optimize both cell yield and quality. Cardiomyocytes are commonly induced in static culture with limited expandability. In this study, we explored the impact of cell adhesion remodeling on hPSC cell fate determination. We reveal that cell passaging at critical time points drives cardiac cell fate even without traditional cardiac inducers. Cardiac fate is specified while cells proliferate continuously. Cell adhesion remodeling leads to a 10-fold increase of cardiomyocyte yield with high purity in comparison to traditional static culture. AMPK activation and PI3K/AKT inhibition were observed following cell passaging. The impact of cell passaging can be mimicked by Src and FAK inhibition, suggesting critical roles of integrin signaling pathway in passaging-driven cardiac differentiation. Transcriptome analysis suggests that cell adhesion remodeling enhances the expression of critical cardiac genes associated with maturation. This study highlights that cell adhesion remodeling significantly impacts cell fate during in vitro differentiation. Our study provides an ideal method for high-yield, high-purity cardiomyocyte production, and offers a useful potential strategy for generating other cell types through directed differentiation.