Fixed Cultivation and Optimization of Dictyostelium discoideum
Abstract：In recent years, the cellular slime mould Dictyostelium discoideum (Dd) has emerged as a promising eukaryotic system for the expression of recombinant pharmaceutical proteins requiring post-translational modifications. However, its application is seriously affected by slow growth rates (doubling time of 8-10 h) as well as low maximal cell densities (1-2×107 mL-1) in the presence of complex axenic medium. For the production of high-value products like heterogenous proteins in substantial quantities it would represent a considerable advantage, if Dd could be cultivated at high cell density.
In consequence, some work have been done here which have led – at least partially – to alleviate these two restrictions. The composition of both complex medium HL-5C and synthetic medium SIH was optimized. A Dictyostelium AX3 strain, AX3-pFH was used expressing the soluble human Fas ligand. we studied the adsorption characteristics of towel carrier in the culture, and optimize a series of parameters in towel immobilization. We find the best way to elute Dictyostelium discoideum cells in the towel carrier is to wash and extrude with 20mL phosphate solution twice.
Key words：Dictyostelium discoideum; mass suspension cultivation; stator
