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Cell Biology
ATCC® Number: CRL-10686™    Price: $407.00
Designations: PG13
Depositors:  Fred Hutchinson Cancer Res. Cntr.
Biosafety Level: 2 [CELLS CONTAIN RETROVIRUS ]
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Mus musculus (mouse)
Morphology: fibroblast
Source: Disease: normal
Strain: NIH/Swiss
Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Applications: transfection host (Roche FuGENE® Transfection Reagents)
Reverse Transcript: positive
Age: embryo
Comments: PG13 is a retrovirus packaging cell line derived from TK- NIH/3T3 cells and based on the Gibbon ape leukemia virus (GaLV).
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 ATCCCRL-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.
Propagation: ATCC 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%.
Temperature: 37.0°C
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
Preservation: Freeze medium: Complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor temperature
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
derivative:ATCC CRL-10685
References: 1818: 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
4473: Miller AD, Rosman GJ. Improved retroviral vectors for gene transfer and expression. BioTechniques 7: 980-990, 1989. PubMed: 2631796
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All prices are listed in U.S. dollars and are subject to change without notice. A discount off the current list price will be applied to most cultures for nonprofit institutions in the United States. Cultures that are ordered as test tubes or flasks will carry an additional laboratory fee. Fees for permits, shipping, and handling may apply.
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