| Applications |
DAN cells were isolated after selection with G418 and screening for helper activity. The line is used as a helper cell for propagating replication incompetent vectors derived from spleen necrosis virus (SNV). After serial passaging for 2 months, the cells appears to decrease in helper activity (as evidence by drops in viral titers produced), thus it is best to prepare generous frozen stocks of the cells upon receipt. |
| Derivation |
DAN cells were isolated after selection with G418 and screening for helper activity. The line is used as a helper cell for propagating replication incompetent vectors derived from spleen necrosis virus (SNV). After serial passaging for 2 months, the cells appears to decrease in helper activity (as evidence by drops in viral titers produced), thus it is best to prepare generous frozen stocks of the cells upon receipt. |
| Comments |
DAN is a retrovirus packaging cell line derive from the D-17 canine osteogenic sarcoma cell line (ATCC CCL-183) by Howard Temin. D-17 cells were transfected with plasmids pBR1 (gag - pol genes from spleen necrosis virus), pJD1 (env gene from amphotropic murine leukemia virus) and pSV2neo (G418 resistance). DAN cells were isolated after selection with G418 and screening for helper activity. The line is used as a helper cell for propagating replication incompetent vectors derived from spleen necrosis virus (SNV). After serial passaging for 2 months, the cells appears to decrease in helper activity (as evidence by drops in viral titers produced), thus it is best to prepare generous frozen stocks of the cells upon receipt. |
| Subculturing |
Protocol: Remove spent medium, add fresh 0.25% trypsin, 0.53 mM EDTA solution, rinse and remove trypsin. Add fresh trypsin solution (1 to 2 ml) and let the culture sit at room temperature (or at 37C) until the cells detach. Add fresh medium, aspirate and dispense into new flasks. Inoculate new flasks with 1 to 2 X 10(5) cells per sq cm. Interval: Subculture at or prior to becoming confluent. Medium Renewal: Twice per week |
| References |
Dougherty JP, et al. New retrovirus helper cells with almost no nucleotide sequence homology to retrovirus vectors. J. Virol. 63: 3209-3212, 1989. PubMed: 2524600
Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.
Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.
Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.
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