Effect of extracellular matrix protein pattern on the differentiation of myoblast

The differentiation mechanism of skeletal myoblast differentiation is still unclear. The main purpose of this research is to explore the effect of the extracellular matrix (ECM) protein pattern on myogenic differentiation. In this study, four different fibronectin micropatterns (parallel, random, vertical, and control group) were prepared by microcontact printing technology, and the differentiation of C2C12 cells after 7 day incubation was analyzed by immunofluorescence staining and analyzes the adhesion molecule, Vinculin and F- actin distribution after 2 h, 6 h, 18 h between groups by immunofluorescence staining method, each group experiment repeated 3 times, each time point had 3 parallel samples. The myotubes number formed in the control group had no significant difference compared with that in the parallel alignment group (17. 33±0. 58 vs 16±1.73), but was significantly higher than the vertical group (7.00 ±1.00) and the random group significantly (10. 89±0. 19) (P<0. 05). At 2 h, the arrangement of microfilaments in cells was consistent with the local micropatterns. At 6 h, the microfilament began to align along the long axis direction of the whole strips pattern. At 12 h, the distribution of microfilaments was consistent with that of the long axis direction of the whole strips pattern. At 18 h, the adhesion plaques formed was distributed along the micropattern area. In conclusion, the extracellular matrix protein patterns are influential on the differentiation of myoblasts, different fibronectin polarity micropatterns have distinct effects on myoblasts cytoskeleton and adhesion, and it might be one of the reasons that later differentiation of C2C12 results in different outcomes.