BALB/3T3 clone A31 Cells

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.

Adherent

BALB/c

2

Supplement the medium with 10% FBS

2 to 3 times per week

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

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.

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.

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

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.

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

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.

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

When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.

We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success.

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