DAN (ATCC® CRL-2130)

Organism: Canis familiaris  /  Disease: osteosarcoma

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Organism Canis familiaris
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2
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 Earle's BSS containing 0.4 mg/ml G418, 92%; fetal bovine serum, 8%
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
Freeze medium: Culture medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Name of Depositor H Temin, DW Burns
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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
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