| Field | Specification |
|---|---|
| Mfr No | |
| Species |
BALB/3T3 clone A31, a fibroblast cell line developed by S.A. Aaronson and G.T. Todaro in 1968, originates from disaggregated 14- to 17-day-old BALB/c mouse embryos. This cell line is a fundamental tool in the study of cellular biology, particularly noted for its capacity to support virus growth and susceptibility to oncogenic transformations. Characteristically, these cells are spindle-shaped fibroblasts that can act as multipotential mesenchymal cells. They demonstrate the potential to differentiate into various tissues depending on microenvironmental influences or culture conditions, underlining their versatility in experimental models.
The cell culture practices for BALB/3T3 clone A31 involve repeated transfers before reaching confluence to minimize cell-cell contact, promoting characteristics such as contact inhibition of cell division, growth at high dilution, and low saturation density. These cells exhibit a karyotype variability with a modal number of 78 chromosomes, ranging from 62 to 109, predominantly featuring telocentric or acrocentric chromosomes. Despite occasional reports of cytogenetic instability, BALB/3T3 A31 cells maintain a non-tumorigenic status, though they show tumorigenic properties when cultured in semisolid mediums. Notably, they are highly susceptible to transformation by oncogenic DNA viruses like SV40 and murine sarcoma virus, and have tested negative for the ectromelia virus (mousepox), adding another layer of value for virological and oncological research.
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Tumorigenic: No, the cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- 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.
- fluidRenewal: 2 to 3 times per week
- 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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