Culturing characteristics of Hanwoo myosatellite cells and C2C12 cells incubated at 37°C and 39°C for cultured meat

J Anim Sci Technol. 2023 May;65(3):664-678. doi: 10.5187/jast.2023.e10. Epub 2023 May 31.

Abstract

To improve culture efficiency of Hanwoo myosatellite cells, these cells were cultured at different temperatures. Hanwoo myosatellite cells were compared with C2C12 cells to observe proliferation and differentiation at culture temperatures of 37°C and 39°C and determine the possibility of using them as cultured meat. Immunofluorescence staining using Pax7 and Hoechst, both cells cultured at 37°C proliferated better than cultured at 39°C (p < 0.05). When differentiated cells were stained with myosin and Hoechst, there was no significant difference in myotube thickness and Fusion index (p > 0.05). In Western blotting analysis, Hanwoo myosatellite cells were no significant difference in the expression of myosin between cells differentiated at the two temperatures (p > 0.05). C2C12 cells were no significant difference in the expression of myosin between cells differentiated at the two temperatures (p > 0.05). In reverse transcription and quantitative polymerase chain reaction (RT-qPCR) analysis, Hanwoo myosatellite cells cultured at 39°C had significantly (p < 0.05) higher expression levels of MyHC, MYF6, and MB than those cultured at 37°C. C2C12 cells cultured at 39°C showed significantly (p < 0.05) higher expression levels of MYOG and MB than those cultured at 37°C. To increase culture efficiency of Hanwoo myosatellite cells, proliferating at 37°C and differentiating at 39°C are appropriate. Since results of temperature differences of Hanwoo myosatellite cells were similar to those of C2C12 cells, they could be used as a reference for producing cultured meat using Hanwoo satellite cells.

Keywords: C2C12 myoblast cell; Culture temperature; Differentiation; Hanwoo myosatellite cell; Proliferation.