Optimal Pre-Plating Method of Chicken Satellite Cells for Cultured Meat Production

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.