Associate Professor Graduate School of Engineering, Osaka University Osaka University, Osaka, Japan
Abstract: To industrialize induced pluripotent stem cell (iPSC)-derived cell therapy, it is necessary to develop robust and highly efficient iPSC differentiation strategies. The cell behavior has been shown to be an important factor regulating cell fate and yield during differentiation. Thus, the spatial variation in cell density during early differentiation may lead to heterogeneity of differentiation. In this study, we explored the concept of “synchronization” in the context of cell behavior-driven mechanical memory in iPSC culture using botulinum hemagglutinin (HA), an E-cadherin function-blocking agent. We showed that temporal disruption of cell-cell interactions by HA results in suppression of spatial heterogeneity of cells within the culture. HA resulted in mechanical memory synchronization with Yes-associated protein (YAP), which increased pluripotent cell homogeneity. Synchronized iPSCs were applied to hepatocyte differentiation medium to test the functional role of synchronized mechanical memory with YAP in regulating cell differentiation. The synchronized iPSCs have higher capability to differentiate into functional hepatocytes than unsynchronized iPSCs, resulting in improved efficiency and robustness of iPSC differentiation. Furthermore, we demonstrated that the synchronization of cell behavior-driven mechanical memory using HA can be used to improve the robustness of differentiation process of various differentiated cell types such as pancreatic progenitor, neuron, and retinal pigment epithelial cells. Our results from the synchronization of mechanical memory using HA confirmed that iPSC differentiation improve the yield and quality of various differentiated cell types, providing a valid tool for various disease modelling and drug discovery applications.
