Oh Sehyuk, Park Sanghun, Park Yunhwan, Kim Yun-A, Park Gyutae, Cui Xiangshun, Kim Kwansuk, Joo Seontea, Hur Sunjin, Kim Gapdon, Choi Jungseok
Department of Animal Science, Chungbuk National University, Cheongju 28644, Korea.
Division of Applied Life Science (BK21 Four), Gyeongsang National University, Jinju 52852, Korea.
J Anim Sci Technol. 2023 May;65(3):664-678. doi: 10.5187/jast.2023.e10. Epub 2023 May 31.
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 ( < 0.05). When differentiated cells were stained with myosin and Hoechst, there was no significant difference in myotube thickness and Fusion index ( > 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 ( > 0.05). C2C12 cells were no significant difference in the expression of myosin between cells differentiated at the two temperatures ( > 0.05). In reverse transcription and quantitative polymerase chain reaction (RT-qPCR) analysis, Hanwoo myosatellite cells cultured at 39°C had significantly ( < 0.05) higher expression levels of MyHC, MYF6, and MB than those cultured at 37°C. C2C12 cells cultured at 39°C showed significantly ( < 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.
为提高韩牛肌卫星细胞的培养效率，将这些细胞在不同温度下培养。将韩牛肌卫星细胞与C2C12细胞进行比较，以观察在37°C和39°C培养温度下的增殖和分化情况，并确定将它们用作培养肉的可能性。使用Pax7和Hoechst进行免疫荧光染色，在37°C培养的两种细胞均比在39°C培养的增殖更好（<0.05）。当用肌球蛋白和Hoechst对分化细胞进行染色时，肌管厚度和融合指数没有显著差异（>0.05）。在蛋白质免疫印迹分析中，在两种温度下分化的韩牛肌卫星细胞之间肌球蛋白的表达没有显著差异（>0.05）。在两种温度下分化的C2C12细胞之间肌球蛋白的表达没有显著差异（>0.05）。在逆转录和定量聚合酶链反应（RT-qPCR）分析中，在39°C培养的韩牛肌卫星细胞中，MyHC、MYF6和MB的表达水平显著高于在37°C培养的细胞（<0.05）。在39°C培养的C2C12细胞中，MYOG和MB的表达水平显著高于在37°C培养的细胞（<0.05）。为提高韩牛肌卫星细胞的培养效率，在37°C增殖并在39°C分化是合适的。由于韩牛肌卫星细胞温度差异的结果与C2C12细胞相似，它们可作为使用韩牛卫星细胞生产培养肉的参考。
