AAV-293

The AAV-293 cell line is a permanent line established from primary embryonic human kidney transformed with human adenovirus type 5 DNA. The genes encoded by the E1 region of adenovirus (E1a and E1b) are expressed in these cells and participate in transactivation of viral promoters, allowing these cells to produce high levels of protein.

AAV-293 is derived from the parental 293 cell line, through cloning and multiple rounds of testing, AAV-293 is specifically selected for a high level of AAV production in a helper-free system. It offers several advantages over the regular 293 cells: Larger cell surface area resulting higher transfection and better yield of AAV.

The advantages are a flattened morphology, firm attachment to culture plate, and the cells are ideal for large-scale culture and AAV production. Adeno-associated virus (AAV) belongs to the family of Parvoviridae, a group of viruses among the smallest of single-stranded and non-enveloped DNA viruses.

There are nine different AAV serotypes reported to date. AAV can infect both dividing and non-dividing cells and can be maintained in the human host cell, creating the potential for long-term gene transfer. Recombinant AAV-2 is the most common serotype used in gene delivery, and can be produced at high titers with a helper virus or AAV-293 cells.

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.