Hollow fiber bioreactor perfusion culture system for magnetic force-based skeletal muscle tissue engineering

Large-scale skeletal muscle tissue cultures are often limited by nutrient supplementation and oxygen diffusion. In the present study, we used a hollow-fiber bioreactor system to supply nutrients and oxygen for the cultivation of high cell-density skeletal muscle tissue constructs fabricated by a magnetic force-based tissue engineering technique. C2C12 cells, magnetically-labeled with magnetite cationic liposomes (MCLs), were mixed with a type I collagen solution and seeded into the cell culture space of the hollow-fiber bioreactor. A magnet was then placed underneath the bioreactor to accumulate MCL-labeled cells in the space between the hollow fibers by magnetic force. Perfusion culture was performed using a myogenic differentiation medium for 7 d. Histological observation revealed that high cell-dense and viable tissue constructs containing myotubes were successfully formed. Furthermore, muscle-specific proteins, such as myosin heavy chain and tropomyosin, were detected by western blot, indicating that C2C12 cells underwent myogenic differentiation. These findings indicate that the hollow-fiber bioreactor system is an effective approach for the in vitro culture of large skeletal muscle tissue constructs, fabricated by magnetic force-based tissue engineering. &copy 2012 The Society of Chemical Engineer, Japan.