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2A (ATCC® CRL-12013™)

Organism: Homo sapiens, human  / 

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Organism Homo sapiens, human
Product Format frozen
Culture Properties adherent
Biosafety Level 2 Cells contain adenoviral DNA sequences

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.
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.
Disclosure This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.
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.
U.S. Patent Number
Disclosure This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC.
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

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
Product Sheet
Product Sheet
Certificate of Analysis
Certificate of Analysis
SDS
SDS
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