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UK Health Security Agency

PA317

PA317

Catalogue No.

89032007

Cell Line Name

PA317

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.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.

Characteristics

Tissue of Origin

Embryo

Karyotype

Not specified

Applications

Transfection and virus studies: herpes virus

Disease

None Stated

Culture Conditions

Cell Type

Fibroblastic

Subculture Routine

Split sub-confluent cultures (70-80%) 1:2 to 1:3 i.e. seeding at 3-5x10,000 cells/cm² using 0.05% trypsin; 5% CO₂; 37°C; Grow in HAT for 5 days followed by 4 days in HT once a month and prior to cryopreservation.Cells should be frozen in 95% cell culture medium (50% Fresh: 50% Conditioned) + 5% DMSO

Culture Medium

DMEM + 2mM Glutamine + 10% Foetal Bovine Serum (FBS).

Growth Mode

Adherent

Additional Info

Depositor

Obtained from ATCC

Country of Origin

United States

GMO Status

Genetically Modified Organism Class 1 (GMO1)

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

Mol. Cell. Biol. 1986;6:2895; N. Engl. J. Med.1990; 323: 570

Bibliography

Not specified

Available Formats

  • Frozen
  • DNA-5µg (100ng/µl)

If use of this culture results in a scientific publication, it should be cited in the publication as: PA317 (ECACC 89032007).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.