Kim So-Hee, Kim Chan-Jin, Lee Eun-Yeong, Son Yu-Min, Hwang Young-Hwa, Joo Seon-Tea
Division of Applied Life Science (BK21 Four), Gyeongsang National University, Jinju 52852, Korea.
Institute of Agriculture & Life Science, Gyeongsang National University, Jinju 52852, Korea.
Food Sci Anim Resour. 2022 Nov;42(6):942-952. doi: 10.5851/kosfa.2022.e61. Epub 2022 Nov 1.
To establish a pre-plating method of chicken satellite cells with high purity, pre-plating was performed under culture conditions of 37°C and 41°C, and the pre-plating time was set from a total of 3 hours to 6 hours in consideration of the cell attachment time. The purity of the cells was confirmed by staining paired box protein 7 (Pax7) after proliferation, and Pax7 expression was the highest in culture flasks shaken for 2 hours after incubation at 41°C for 2 hours to prevent the attachment of satellite cells (p<0.05). Also, when pre-plating and proliferation were performed at 37°C and 41°C, the Pax7 expression rate was higher at 41°C. The differentiation capabilities of the three groups (T3, T6, and T7) with high Pax7 expression were compared and the fusion index (%) and myotube formation area (%) determined by myosin heavy chain (MHC) staining was calculated. The T6 and T7 groups, which were cultured at 41°C, showed significantly higher values than the T3 group (p<0.05). There was no significant difference in the expression of Pax7 and MHC between the T6 and T7 groups (p>0.05). These results suggest that pre-plating at 41°C for a total of 4 hours was the most efficient in terms of cost and time for purifying chicken satellite cells for cultured meat.
为建立一种高纯度鸡卫星细胞的预铺板方法,在37°C和41°C的培养条件下进行预铺板,并考虑细胞贴壁时间将预铺板时间设置为总共3小时至6小时。增殖后通过配对盒蛋白7(Pax7)染色确认细胞纯度,在41°C孵育2小时后振荡培养瓶2小时以防止卫星细胞贴壁,此时Pax7表达最高(p<0.05)。此外,当在37°C和41°C进行预铺板和增殖时,41°C时Pax7表达率更高。比较了Pax7高表达的三组(T3、T6和T7)的分化能力,并计算了通过肌球蛋白重链(MHC)染色测定的融合指数(%)和肌管形成面积(%)。在41°C培养的T6和T7组的值显著高于T3组(p<0.05)。T6和T7组之间Pax7和MHC的表达没有显著差异(p>0.05)。这些结果表明,就成本和时间而言,在41°C预铺板总共4小时对于纯化用于培养肉的鸡卫星细胞是最有效的。