Cell Line Origin
Mouse fibroblast with herpes TK gene
Cell Line Description
The cell line PA317 was generated by co-transfecting NIH 3T3 TK- cells with retrovirus packaging construct DNA (pPAM3) and the herpes simplex virus thymidine kinase (TK) gene carried as a fragment in pBR322. The construct pPAM3 contains several mutations in addition to deletion of the packaging signal. These features appear to prevent the production of helper virus and the transfer of packaging function. Transfection or infection with retroviral vectors results in the production of retrovirus virions with an amphotropic host range. These are capable of infecting mammalian cells and have been applied to transfer genes into humans. PA317 cells will loose their packaging ability during long-term culture. Therefore, a selection should be included routinely and before cryopreservation of cells. The selection procedure includes growth in HAT-containing medium (0.03mM hypoxanthine, 0.001mM amethopterin, 0.02mM thymidine) for 5 days. To dilute residual amethopterin cells should then be cultured for 4 days in HT containing medium (0.03mM hypoxanthine, 0.02mM thymidine). Cells are stable for at least 1 month after selection.
Application
Transfection and virus studies: herpes virus
Culture Medium
DMEM + 2mM Glutamine + 10% Foetal Bovine Serum (FBS).
Subculture Routine
Split sub-confluent cultures (70-80%) 1:2 to 1:3 i.e. seeding at 3-5x10,000 cells/cm2 using 0.25% trypsin; 5% CO2; 37°C; Grow in HAT for 5 days followed by 4 days in HT once a month and prior to cryopreservation
Other Notes
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biological source: mouse embryo. growth mode: Adherent. karyotype: Not specified. morphology: Fibroblast. products: Not specified. receptors: Not specified. technique(s): cell culture | mammalian: suitable. shipped in: dry ice. storage temp.: −. 196°C.- UPC:
- 41116133
- Condition:
- New
- Weight:
- 1.00 Ounces
- HazmatClass:
- No
- WeightUOM:
- LB
- MPN:
- 89032007-1VL
