PA317

NOTE: Both the cell line and DNA from the cell line may be available for this product. Please choose -1VL or VIAL for cells, or -DNA-5UG for DNA.

• UNSPSC Code: 41106514

Properties

• biological source: mouse embryo
• packaging: tube of 5 μg 89032007-DNA-5UG; pkg of vial of cells 89032007-1VL
• 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

Description

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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