| Field | Specification |
|---|---|
| Species |
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.
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- Isoenzymes: G6PD, B
- Viruses: Contains Papovavirus
- Virus susceptibility: Herpes simplex, vesicular stomatitis (Indiana), poliovirus 2
- Reverse transcriptase: Negative
- Karyotype: Hyperdiploid, Modal number: 73-78
- cultureMedium: EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- seedingDensity: 1 x 104 cells/cm2
- fluidRenewal: 1 to 2 times per week
- postThawRecovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 48 hours.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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