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Age:
embryo
Comments:
Initial cultures from a thawed ampule may take 10 to 12 days to become confluent. Tested and found negative for ectromelia virus (mousepox).
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing:
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:6 is recommended Medium Renewal: 2 to 3 times per week Remove medium, add fresh 0.25% trypsin, 0.03% EDTA for 1 to 3 minutes, remove trypsin and let the culture sit at 37C for 10 to 20 minutes. Add fresh medium, aspirate and dispense into new flasks.
References:
22696: Hartley JW, Rowe WP. Clonal cells lines from a feral mouse embryo which lack host-range restrictions for murine leukemia viruses. Virology 65: 128-134, 1975. PubMed: 167514