2A (ATCC® CRL-12013)

Organism: Homo sapiens, human  / 

Organism Homo sapiens, human
Product Format frozen
Culture Properties adherent
Biosafety Level 2 Cells contain adenoviral DNA sequences
Age fetus
Applications
The 2A amphotropic retroviral packaging cell line was derived from the human embryonic kidney line, 293 (see ATCC CRL-1573).
After selection for transfected cells, individual drug resistant cell colonies were expanded, analyzed and selected for overexpression of MoMLV gag/pol.
Derivation
The 2A amphotropic retroviral packaging cell line was derived from the human embryonic kidney line, 293 (see ATCC CRL-1573).
Comments
The 2A amphotropic retroviral packaging cell line was derived from the human embryonic kidney line, 293 (see ATCC CRL-1573).
293 cells were co-transfected with the methotrexate resistance vector, pFR400 and the MoMLV gag/pol expression vector, pSCV10.
After selection for transfected cells, individual drug resistant cell colonies were expanded, analyzed and selected for overexpression of MoMLV gag/pol.
293 (clone 2-3) was co-transfected with the phleomycin resistance vector, pUT507 and the amphotropic envelope expression vector, pMLPenvAmSph and subsequently cloned.
The 2A clone was selected for expression of relatively high levels of both gag/pol and amphotropic envelope proteins.
Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:
  • O.1 mM Non-Essential Amino Acids (NEAA)
  • fetal bovine serum to a final concentration of 10%

Subculturing
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days
Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37C) until the cells detach.
Add fresh culture medium, aspirate and dispense into new culture flasks.
Cryopreservation
culture medium 95%; DMSO, 5%
Culture Conditions
Temperature: 37.0°C
Name of Depositor Chiron Viagene, Inc.
References

DePolo NJ, et al. The resistance of retroviral vectors produced from human cells to serum inactivation in vivo and in vitro is primate species dependent. J. Virol. 73: 6708-6714, 1999. PubMed: 10400768

Barber JR, et al. Retroviral packaging cell lines. US Patent 5,591,624 dated Jan 7 1997

Basic Documentation
References

DePolo NJ, et al. The resistance of retroviral vectors produced from human cells to serum inactivation in vivo and in vitro is primate species dependent. J. Virol. 73: 6708-6714, 1999. PubMed: 10400768

Barber JR, et al. Retroviral packaging cell lines. US Patent 5,591,624 dated Jan 7 1997