An attempt for improving plating efficiency of human diploid fibroblasts

An attempt was made to enhance the plating efficiency of normal human diploid skin fibroblasts by testing various culture media. Eagle's minimum essential medium (MEM), Dulbecco's MEM, William's medium E, DM-160, McCoy's 5a, RPMI-1640, Ham's F12, MCDB 102, Waymouth's MB 752/1, and L-15 synthetic media were examined. L-15 was the best among them. Plating efficiency with L-15 supplemented with 20% calf serum ranged from 20 to 60%. Plating efficiency decreased in the order of Eagle's MEM, Dulbecco's MEM, DM-160, and Ham's F12. Waymouth's MB 752/1, MCDB 102 McCoy's 5a, RPMI-1640, and William's medium E were not favorable for high plating efficiency. Although L-15 gave the best plating efficiency and Waymouth's MB 752/1 the worst, the Waymouth medium supported better cell proliferation than L-15, when cell proliferation was compared in mass culture with these two media. It was concluded that L-15 was better suited for culture of small number of cells, and especially for work involving colony formation.