Effects of culture temperature (37°C, 39°C) and oxygen concentration (20%, 2%) on proliferation and differentiation of C2C12 cells

Cells, culture media, and so on are important elements of cultured meat production technology. Also, the environment (humidity, temperature, air composition, dissolved oxygen tension, etc.) for in vivo muscle production are important. Among cell culture conditions, culture temperature and oxygen concentration are important physical factors that can affect cells. The objective of this study was to determine effects of culture temperature and oxygen concentration on proliferation and differentiation of muscle cells. This study was conducted using C2C12 cells of rat myoblasts widely used in muscle physiology. The temperature was chosen to induce some thermal stress at 39°C, and the oxygen concentration was selected at 2% to mimic the oxygen levels present in muscle cells in vivo. Culture conditions consisted of CON (37°C/20% O2), T1 (37°C/2% O2), T2 (39°C/20% O2) and T3 (39°C/2% O2). In terms of cell proliferation, temperature conditions had a significant impact (p < 0.05), and a temperature of 39°C was found to reduce the cell count. Oxygen conditions had a significant impact on 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) absorbance, and temperature conditions were found to have a greater influence on cell proliferation compared to oxygen condition and interaction condition. The temperature conditions were found to have a significant impact (p < 0.05) on the expression of proteins related to myogenesis compared to oxygen conditions. The significant increase (p < 0.05) in the protein expression levels of Myh, Myod1, Myog, and Mb in T2 compared to CON, and in T3 compared to T1, suggests that a temperature of 39°C enhances the expression of myogenic differentiation proteins. These results indicate that temperature conditions have a significant impact (p < 0.05) on cell proliferation and differentiation, more so than oxygen conditions and interaction conditions. And a temperature of 39°C was found to inhibit cell proliferation, but in the case of differentiation, it was observed to be promoted due to the upregulation of myogenic differentiation proteins.