Park Gyutae, Park Sanghun, Oh Sehyuk, Choi Nayoung, Choi Jungseok
Department of Animal Science, Chungbuk National University, Cheongju 28644, Korea.
J Anim Sci Technol. 2025 Jan;67(1):224-235. doi: 10.5187/jast.2023.e130. Epub 2025 Jan 31.
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 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% O), T1 (37°C/2% O), T2 (39°C/20% O) and T3 (39°C/2% O). In terms of cell proliferation, temperature conditions had a significant impact ( < 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 ( < 0.05) on the expression of proteins related to myogenesis compared to oxygen conditions. The significant increase ( < 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 ( < 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.
细胞、培养基等是培养肉生产技术的重要要素。此外，肌肉生产的环境（湿度、温度、空气成分、溶解氧张力等）也很重要。在细胞培养条件中，培养温度和氧气浓度是会影响细胞的重要物理因素。本研究的目的是确定培养温度和氧气浓度对肌肉细胞增殖和分化的影响。本研究使用了在肌肉生理学中广泛应用的大鼠成肌细胞C2C12细胞进行。温度选择为39°C以诱导一定的热应激，氧气浓度选择为2%以模拟体内肌肉细胞中的氧气水平。培养条件包括CON（37°C/20% O）、T1（37°C/2% O）、T2（39°C/20% O）和T3（39°C/2% O）。在细胞增殖方面，温度条件有显著影响（<0.05），发现39°C的温度会减少细胞数量。氧气条件对3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲氧基苯基)-2-(4-磺基苯基)-2H-四唑鎓（MTS）吸光度有显著影响，并且发现温度条件相比于氧气条件和相互作用条件对细胞增殖的影响更大。与氧气条件相比，发现温度条件对与肌生成相关的蛋白质表达有显著影响（<0.05）。与CON相比，T2中Myh、Myod1、Myog和Mb的蛋白质表达水平显著增加（<0.05），与T1相比，T3中也是如此，这表明39°C的温度会增强肌源性分化蛋白的表达。这些结果表明，温度条件对细胞增殖和分化有显著影响（<0.05），比氧气条件和相互作用条件的影响更大。并且发现39°C的温度会抑制细胞增殖，但在分化的情况下，观察到由于肌源性分化蛋白的上调而促进了分化。
