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Three‐dimensional co‐culture of C2C12/PC12 cells improves skeletal muscle tissue formation and function
Authors:Serge Ostrovidov  Samad Ahadian  Javier Ramon‐Azcon  Vahid Hosseini  Toshinori Fujie  S. Prakash Parthiban  Hitoshi Shiku  Tomokazu Matsue  Hirokazu Kaji  Murugan Ramalingam  Hojae Bae  Ali Khademhosseini
Affiliation:1. Advanced Institute for Materials Research (WPI), Tohoku University, Sendai, Japan;2. Laboratory of Applied Mechanobiology, Department of Health Sciences and Technology, ETH, Zurich, Switzerland;3. Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda University, Tokyo, Japan;4. Graduate School of Environmental Studies, Tohoku University, Sendai, Japan;5. Department of Bioengineering and Robotics, Graduate School of Engineering, Tohoku University, Sendai, Japan;6. Centre for Stem Cell Research, A unit of the Institute for Stem Cell Biology and Regenerative Medicine, Christian Medical College Campus, Vellore, India;7. Institut National de la Santé et de la Recherche Médicale U977, Faculté de Chirurgie Dentaire, Université de Strasbourg, France;8. College of Animal Bioscience and Technology, Department of Bioindustrial Technologies, Konkuk University, Seoul, Republic of Korea;9. Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul, Republic of Korea;10. Biomaterials Innovation Research Center, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School and Harvard–MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA;11. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA;12. Center of Nanotechnology, King Abdulaziz University, Jeddah, Saudi Arabia
Abstract:Engineered muscle tissues demonstrate properties far from native muscle tissue. Therefore, fabrication of muscle tissues with enhanced functionalities is required to enable their use in various applications. To improve the formation of mature muscle tissues with higher functionalities, we co‐cultured C2C12 myoblasts and PC12 neural cells. While alignment of the myoblasts was obtained by culturing the cells in micropatterned methacrylated gelatin (GelMA) hydrogels, we studied the effects of the neural cells (PC12) on the formation and maturation of muscle tissues. Myoblasts cultured in the presence of neural cells showed improved differentiation, with enhanced myotube formation. Myotube alignment, length and coverage area were increased. In addition, the mRNA expression of muscle differentiation markers (Myf‐5, myogenin, Mefc2, MLP), muscle maturation markers (MHC‐IId/x, MHC‐IIa, MHC‐IIb, MHC‐pn, α‐actinin, sarcomeric actinin) and the neuromuscular markers (AChE, AChR‐ε) were also upregulated. All these observations were amplified after further muscle tissue maturation under electrical stimulation. Our data suggest a synergistic effect on the C2C12 differentiation induced by PC12 cells, which could be useful for creating improved muscle tissue. Copyright © 2014 John Wiley & Sons, Ltd.
Keywords:muscle  C2C12  PC12  methacrylated gelatin  hydrogel  myotubes  myogenesis
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