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微信小章| 产品名称： | DAN |
|---|---|
| 商品货号： | TS212281 |
| Organism： | Canis familiaris, dog |
| Tissue： | bone |
| Product Format： | frozen |
| Morphology： | fibroblast |
| Culture Properties： | adherent |
| Biosafety Level： | 2
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease： | osteosarcoma |
| Age： | 11 years |
| Gender： | female |
| Strain： | poodle |
| 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. |
| Storage Conditions： | liquid nitrogen vapor phase |
| 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. |
| Clinical Data： | female |
| 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. |
| Complete Growth Medium： | Minimum essential medium (Eagle) with Earles BSS containing 0.4 mg/ml G418, 92%; fetal bovine serum, 8%
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| 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 |
| Cryopreservation： | Complete growth medium described above supplemented with 5% (v/v) DMSO.xa0 Cell culture tested DMSO is available as ATCC Catalog No. 4-X. |
| Culture Conditions： | Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5% |
| Name of Depositor： | H Temin, DW Burns |
| Deposited As： | Canis familiaris |
| 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. |