Suspension culture of an antibacterial peptide producing cell line from Bombina orientalis

The suspension culture of an anchorage-dependent cell line (Bok-1) from Bombina orientalis was successful in respects of cost and efficiency. The amount of cells obtained from the suspension culture was almost equivalent to that from the anchorage-dependent culture. This result shows the possibility of suspension culture for scale-up. The cells in suspension produced an antibacterial peptide as much as anchorage-dependent cells did. The cell growth (6.0 x 10⁶ cells/ml) and viability (>80%) at 10 rpm were higher than that at 0 rpm (1.9 x 10⁶ cells/ml, 65~80%) and 30 rpm (1.8 x 10⁶ cells/ml, 40~76%). The size of cells became smaller at the agitation rate of 30 rpm. The antibacterial activities of cell extracts from suspension cultured cells were confirmed against gram-negative and gram-positive bacteria by the inhibition zone assay and the liquid growth inhibition assay.