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. Cells supplied by HPACC will have been grown in non-selective medium for approximately 2 weeks prior to cryopreservation. Therefore the cells should be stable for at least 2 weeks before requiring a repeat of the selection procedure previously described.
Adherent Suspension
Adherent
Suggested Medium
DMEM + 2mM Glutamine + 10% Foetal Bovine Serum (FBS).
Quality Control
Cell lines were tested and found to be free of mycoplasma, bacterial, viruses, and other toxins. All cells were above 95% viability before freezing.
Shippment
The cells are shipped using dry ice.
Storage
Store the cell lines in liquid nitrogen vapor (less than -130°C)
Warranty
Our cell products are for research use only. They are not authorized for human, in vitro diagnostic procedures or for therapeutic procedures.
