Spatial cell fate manipulation of human pluripotent stem cells by controlling the microenvironment using photocurable hydrogel

Human pluripotent stem cells (hPSCs) sense the surrounding chemical, and physical microenvironment and adjust their behavior accordingly. However, current in vitro research is mostly conducted in plastic culture wares which provides an oversimplified microenvironment compared to that in vivo. Increasing evidence has highlighted the importance of the physical microenvironment in hPSCs differentiation and maintenance. Here, hPSCs were cultured on homogeneous substrates with specific stiffness. Biocompatible material that allows spatial control of the chemical and mechanical environment for hPSC culture has not yet been developed. Culture substrates that allow flexible manipulation of spatial physical properties can assist the recapitulation of in vivo environment, and assist tissue engineering. We developed a photocurable polyethylene glycol–polyvinyl alcohol (PVA-PEG) hydrogel which allows spatial control of the surface stiffness and structure at a micrometer level. The developed hydrogel can be functionalized using different extracellular matrix (ECM) proteins. Laminin 511 functionalized PVA-PEG gel supports hPSCs growth and differentiation. Furthermore, by spatially controlling the stiffness of the patterned gel, we could selectively differentiate cells, generating complex patterned structures.