Human pluripotent stem cell (hPSC-) derived cardiomyocytes have potential applications in drug discovery, toxicity testing, developmental studies, and regenerative medicine. Before these cells can be reliably utilized, characterization of their functionality is required to establish their similarity to native cardiomyocytes. We tracked fluorescent beads embedded in 4.4–99.7 kPa polyacrylamide hydrogels beneath contracting neonatal rat cardiomyocytes and cardiomyocytes generated from hPSCs via growth-factor-induced directed differentiation to measure contractile output in response to changes in substrate mechanics. Contraction stress was determined using traction force microscopy, and morphology was characterized by immunocytochemistry for α-actinin and subsequent image analysis. We found that contraction stress of all types of cardiomyocytes increased with substrate stiffness. This effect was not linked to beating rate or morphology. We demonstrated that hPSC-derived cardiomyocyte contractility responded appropriately to isoprenaline and remained stable in culture over a period of 2 months. This study demonstrates that hPSC-derived cardiomyocytes have appropriate functional responses to substrate stiffness and to a pharmaceutical agent, which motivates their use in further applications such as drug evaluation and cardiac therapies.

Inspired by natural examples of swelling-actuated self-folding, we utilize photodegradable hydrogels as dynamically tunable, shape-changing scaffolds for culturing cells. Poly(ethylene glycol) diacrylate-based thin films incorporating ortho-nitrobenzyl (o-NB) moieties are transformed from flat 2D sheets to folded 3D structures by exposure to 365 nm UV light. As the UV light is attenuated through the thickness of the gel, a cross-link density gradient is formed. This gradient gives rise to differential swelling and a bending moment, resulting in gel folding. By tuning the UV light dose and the molar ratio of photodegradable to nondegradable species, both the initial degree of folding and the relaxation of tubular structures can be accurately controlled. These self-folding photodegradable gels were further functionalized with a cell-adhesive RGD peptide for both seeding and encapsulation of C2C12 mouse myoblasts. Light-induced folding of RGD functionalized hydrogels from flat sheets to tubular structures was demonstrated 1 or 3 days after C2C12 seeding. The C2C12s remained adhered on the inner walls of folded tubes for up to 6 days after folding. The minimum measured diameter of a tubular structure containing C2C12s was 1 mm, which is similar to the size of muscle fascicles. Furthermore, the viability of encapsulated C2C12s was not adversely affected by the UV light-induced folding. This is the first account of a self-folding material system that allows 2D-3D shape change in the presence of both seeded and encapsulated cells at a user-directed time point of choice.

Highly conjugated, hydrophobically modified gelatin hydrogels were synthesized, polymerized and degraded with orthogonal wavelengths of light.

Hydrogels are attractive materials for generating 4D shapes due to their ability to undergo pronounced volume changes in response to several stimuli, including light. We previously reported shape-changing hydrogels actuated by longwave UV and visible light in the presence of live cells using poly(ethylene glycol) macromers incorporating different photodegradable ortho-nitrobenzyl (o-NB) groups. In this comprehensive study, we determine the effect of chemical structure of different o-NB macromers (which influences molar absorptivity and rate constant of degradation), composition (macromer weight percent), fabrication design (initial gel thickness) and environment (ionic strength of solution) on light-induced hydrogel folding. We demonstrate successful photopolymerization and subsequent photodegradation of hydrogels, multistep folding, and live-cell encapsulation. This hydrogel system may be useful as new tool in stem cell differentiation and developmental biology research, facilitating the in vitro investigation of processes that are sensitive to both physical and temporal stimuli.