We deviced a new method for measurement of cell propagation rate which was more easy and accurate than the conventional "replicate culture method", and discussed on some experimental conditions. At first, the effect of cellular concentration on propagation was studied to determine the optimum concentration at seeding. When MA-30 culture flask was used, the optimum one was 3.2 × 10^4 cells/ml. Then, the effect of medium components was studied and it became apparent that RFL cells could divide at least one time and propagate for 50 hours even in the serum-free synthesized medium. On the other hand, the cells could not propagate but begin to liberate from the glass wall of the culture flask after 6 hour-incubation in a balanced salts solution, PBS. Thus the period of PBS treatment should be restricted within 6 hours. Furthermore, cellular toxicity of dimethyl sulfoxide, DMSO, was tested to know its permissible concentration. Since the treatment with 1 % DMSO for 6 hours had little effect on the propagation of RFL cells, this reagent may be a good solvent in addition tests of water-insoluble substances.
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