PG13 (ATCC® CRL-10686)

Organism: Mus musculus, mouse  /  Disease: Leukemia

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Organism Mus musculus, mouse
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2 CELLS CONTAIN RETROVIRUS

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 Leukemia
Age embryo
Strain NIH/Swiss
Applications
transfection host
Storage Conditions liquid nitrogen vapor temperature
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
PG13 is a retrovirus packaging cell line derived from TK- NIH/3T3 cells and based on the Gibbon ape leukemia virus (GaLV).
Comments

Introduction of retroviral vectors by infection or transfection results in the production of retrovirus virions capable of infecting cells from many species (with the exception of mice).

Selection against loss of the DNAs conferring the packaging functions can be performed by growing the cells in medium containing dialyzed fetal bovine serum and 100 nM amethopterin for 5 days followed by cultivation in medium containing HAT and untreated fetal bovine serum for an additional 5 days.After selection, the cells should be maintained in medium containing HT for 2 days to avoid toxic effects due to residual amethopterin.

A derivative of this line is available (PG13/LN c8, see ATCC CRL-10685) that produces a neomycin resistance transducing vector.

Although virus production has not been observed, there is a possibility that the cells will produce a virus similar to GaLV (a moderate risk oncogenic virus) and Biosafety Level 2 or higher precautions should be taken when using this cell line.

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: fetal bovine serum to a final concentration of 10%.
Subculturing
Protocol: Remove medium, add fresh 0.25% Trypsin, 0.53 mM EDTA solution and allow the flask to sit at room temperature (or 37C) until the cells detach (2 to 3 minutes). Add fresh medium, aspirate and dispense into new flasks.
Subcultivation Ratio: A subcultivation ratio of 1:8 is recommended
Medium Renewal: Every 2 to 3 days
Cryopreservation
Freeze medium: Complete growth medium, 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions
Temperature: 37°C
Name of Depositor Fred Hutchinson Cancer Res. Cntr.
Deposited As Mus musculus
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

Miller AD, et al. Construction and properties of retrovirus packaging cells based on gibbon ape leukemia virus. J. Virol. 65: 2220-2224, 1991. PubMed: 1850008

Miller AD, Rosman GJ. Improved retroviral vectors for gene transfer and expression. BioTechniques 7: 980-990, 1989. PubMed: 2631796

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.

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

Miller AD, et al. Construction and properties of retrovirus packaging cells based on gibbon ape leukemia virus. J. Virol. 65: 2220-2224, 1991. PubMed: 1850008

Miller AD, Rosman GJ. Improved retroviral vectors for gene transfer and expression. BioTechniques 7: 980-990, 1989. PubMed: 2631796

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.