WI 38 VA13 subline 2RA

The WI-38 VA13 subline 2RA, derived from the historic WI-38 cell line originally sourced from the lung tissue of a 3-month-old fetus, represents a key advancement in cell culture technology. The original WI-38 cell line was crucial in developing vaccines for numerous viral diseases, such as measles, mumps, rubella, and hepatitis A. The VA13 subline 2RA is an immortalized variant of this cell line, achieved through transformation with Simian Virus 40 (SV40), a practice common in the development of immortal cell lines which allows for indefinite cell replication beyond the standard senescence point of about 50 population doublings.

The incorporation of SV40 into the WI-38 cells to create the VA13 subline 2RA extends the cells' lifespan, providing a more durable model for long-term experiments. This transformation maintains the fundamental properties of the original diploid cells but alters their lifecycle and growth patterns, enabling sustained growth and facilitating extensive studies that were not possible with the finite lifespan of the parent cell line. This makes the VA13 subline particularly useful in ongoing and extensive research areas, including virology, pharmacology, and genetic research, where extended observation periods are necessary.

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.