コイ仔魚由来の新しい細胞系について

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An attempt was made to obtain the primary cell line from carp sac-fry (Cyprinus carpio) with high growth rate and without contamination. Eggs just before hatching were incubated in still-water under aeration in a glass vessel. To avoid contamination by fungi and bacteria, malachite green and nifurprazine-HCI were added in a low concentration to the incubating water. The sac-fries just hatched were pooled in a petri dish and washed several times with Earle's BSS contained penicillin (250 units/ml), streptomycm (250μg/ml), kanamycin(25μg/ml) and fungizone (125μg/ml). After choppfg the fries into small pieces cells were dispersed with pronase-P (0.01%in Eagle's MEM) at room temperature. The cells at a concentration of 1-1.5×107 cells/ml were cultivated in Eagle's MEM with 10% fetal bovine serum (GM-10) at 30°C. For subcultivation primary cells grown to confluent monolayers were dispersed by EDTA-trypsin solution at room temperature. After 3 to 4 days cell sheets of mixed epithelium-like and fibroblast-like cells became con-fluent. As time passed, epithelium-like cells became dominant. The cells grew at 15°C to 37°C. At 30°C the cells could be subcultured at 4 to 7 day intervals. The cells maintained at 15°C could be subcultured at 2 to 3 month intervals. The cells grew well in Eagle's MEM with 5% calf serum as well as in GM-10. At a concentration of 3-4×1.06 cells/ml, the cells in GM-10 added 10% DMSO could be presorved by freezing at -80°C for at least 1 month. The cells could not support the replication of IPN virus. This primary cell designated as CSF cell has been subcultured 42 times over a period of 24 months.
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コイ仔魚由来の新しい細胞系について

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