Effect of cell-extracellular matrix interaction on myogenic characteristics and artificial skeletal muscle tissue
Section snippets
Cell culture
C2C12 cells (within 10 passages; ATCC, Manassas, VA, USA) were seeded at the density of 2000 cells/cm2, and cultured in low glucose Dulbecco's modified Eagle's medium (DMEM; Nacalai Tesque, Kyoto, Japan) with 10% fetal bovine serum (FBS; Equitech-Bio, Kerrville, TX, USA), Penicillin-Streptomycin Mixed Solution (Nacalai Tesque), and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES; Dojindo, Kumamoto, Japan) at 37°C in a 5% CO2 incubator. To induce myogenic differentiation, growth medium
Effect of ECM proteins on migration, proliferation, differentiation, and fusion of C2C12 cells
To investigate the effects of different ECM proteins on C2C12 cells the C2C12 myoblasts were cultured on the fibronectin, laminin, collagen I, collagen IV, and Matrigel coated culture surface. Time-lapse analysis showed that the C2C12 cells on each ECM coated surface migrated with significantly higher speed than that of cells on the control tissue culture surface (Fig. 1A). Next, to examine the effect of ECM protein on the cell viability of C2C12 cells, we cultured the cells in differentiation
Discussion
In this study, we demonstrated that the physical force generation of artificial skeletal muscle tissues could be improved using SU9516 and Matrigel, which contribute to the creation of a suitable myogenic environment in vitro. For drug screening and testing, the use of established cell lines, such as mouse C2C12 cells provide reproducible and repeatable systems, which resulted in the reduction of animal studies. Furthermore, drug screening using artificial skeletal muscle tissue would
Acknowledgments
This work was supported in part by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion Science, Japan (Nos. 18K14063). We thank Kyoto University Radioisotope Research Center for use of facilities.
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Much of this work forms the basis of the Ph.D. dissertation of Ran Ding.