PA317

PA317 is a murine fibroblast cell line that has been engineered to stably express an amphotropic retroviral packaging system. This cell line is widely used in gene therapy and molecular biology research as a producer of recombinant retroviruses capable of transducing a broad range of mammalian cells. PA317 is derived from NIH/3T3 fibroblasts and retains a fibroblast morphology while exhibiting the ability to produce high-titer retroviruses.

PA317 cells are designed to provide essential retroviral components in trans, including gag, pol, and env genes, while lacking replication-competent virus sequences. This ensures safety in experimental and therapeutic contexts by preventing the formation of replication-competent retroviruses. The amphotropic envelope protein expressed by PA317 cells enables the produced retroviruses to infect a diverse array of cell types across different species, making this cell line particularly valuable for applications requiring efficient gene transfer.

This cell line is commonly employed to produce retroviral vectors for gene delivery in preclinical studies, including cancer research, genetic engineering, and regenerative medicine. It is also a preferred choice for validating retroviral vector constructs and optimizing viral production protocols. PA317 provides a reliable platform for retrovirus-mediated gene transfer, supporting advances in gene therapy and functional genomics research.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.