We examined the effect of substrate stiffness on the beating rate, force of contraction, and cytoskeletal structure of embryonic chicken cardiac myocytes by culturing them on laminin-coated polyacrylamide (PA) substrates. Cells cultured on PA substrates with elasticity comparable to that of the native myocardium (18 kPa) exhibited the highest beating rate during the first few days of culture. The initial beating rate of individual cells on all the substrates varied significantly but began to converge within 5 days. We also examined the focal adhesions (FAs) and cytoskeletal structure on different substrates via confocal microscopy and found a higher percentage of FAs on tissue culture (TC) plastic dishes compared with the softer PA gels. Furthermore, highly aligned sarcomeric striations were clearly visible on 18 kPa, 50 kPa, and TC dish, whereas cells on 1 kPa only showed nonaligned diffused striations. The force of contraction on these substrates was measured using a micro-electromechanical system force sensor, which showed that the force of contraction for the cells on TC dishes (F = 71.30 ± 6.38 nN) was significantly larger than those cultured on the 18-kPa PA gel (F = 30.16 ± 3.83 nN). This is most likely due to the formation of higher percentage of FAs on the TC dishes compared with fewer FAs on the softer gels. Our cumulative findings can have a significant impact on the design of 3D cardiac tissue engineered scaffolds.
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