Components of extracellular matrix significantly influence the growth and differentiation of the extracellular matrix. The aim of experiments in vitro is to obtain a mature form of muscle fibers which most closely resemble the state in vivo. Therefore, substrates such as Matrigel and Geltrex, which mimic the composition and properties of the extracellular matrix, are frequently used. Our aim was to compare growth and differentiation markers of primary human skeletal muscle cells in wells without coating and in wells coated with Matrigel or Geltrex. Using Western blot, we determined the abundance of desmin, myosin heavy chain, α1, α2 and β1 subunits of Na+/K+ ATPase (ATP1A1, ATP1A2, ATP1B1) and cleaved caspase 3. The abundance of desmin, MYH7 and α1 subunit of Na+/K+ ATPase were significantly higher in cells grown on Matrigel or Geltrex suggesting enhanced growth and differentiation. Abundance of cleaved caspase 3 seemed to be dependent on the density of cells and not the culturing conditions. Protein concentration measurements showed significantly increased concentration in coated wells. Using RT qPCR with peptidylpropyl isomerase A as an endogenous control, we determined that the mRNA expression levels of MYH7, desmin and Na+/K+ ATPase did not significantly differ between the groups, however, the total mRNA was higher in coated wells. Our results indicate that Matrigel and Geltrex positively influence the differentiation of primary human skeletal muscle cells and provide a basis for further research into the optimization of in vitro models of skeletal muscle.
|
