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Effects of heat stimulation and l‐ascorbic acid 2‐phosphate supplementation on myogenic differentiation of artificial skeletal muscle tissue constructs

Although skeletal muscle tissue engineering has been extensively studied, the physical forces produced by tissue‐engineered skeletal muscles remain to be improved for potential clinical utility. In this study, we examined the effects of mild heat stimulation and supplementation of a l‐ascorbic acid derivative, l‐ascorbic acid 2‐phosphate (AscP), on myoblast differentiation and physical force generation of tissue‐engineered skeletal muscles. Compared with control cultures at 37°C, mouse C2C12 myoblast cells cultured at 39°C enhanced myotube diameter (skeletal muscle hypertrophy), whereas mild heat stimulation did not promote myotube formation (differentiation rate). Conversely, AscP supplementation resulted in an increased differentiation rate but did not induce skeletal muscle hypertrophy. Following combined treatment with mild heat stimulation and AscP supplementation, both skeletal muscle hypertrophy and differentiation rate were enhanced. Moreover, the active tension produced by the tissue‐engineered skeletal muscles was improved following combined treatment. These findings indicate that tissue culture using mild heat stimulation and AscP supplementation is a promising approach to enhance the function of tissue‐engineered skeletal muscles. Copyright © 2015 John Wiley & Sons, Ltd.