Description: AB12 is a mouse mesothelioma cell line. Mice (BALB/c strain, female, 6-8 weeks old) were exposed to crocidolite asbestos through intraperitoneal injection, resulting in tumour development. Cultures were established from malignant mesothelial cells obtained from ascites fluid. Cells were cultured successfully for at least 9 months and 32 passages. 

Organism: Mouse (Mus musculus) 

Strain: BALB/c

Tissue: Mesothelium; malignant mesothelial cells were obtained from ascites fluid.

Growth Properties: Adherent

Morphology: Cells are stellate shaped.

Growth Medium: RPMI 1640 (with 2mM L-Glutamine + 25mM HEPES) + 5% Foetal Calf Serum 

Resuscitation: Remove protective cryoflex layer around the ampoule prior to thawing. Thaw the ampoule by gently agitating in a 37°C waterbath; thawing should be rapid (around 2 minutes). A centrifugation step to remove the cryoprotectant after thawing is necessary for this cell line. Recovery from thaw may take 3 days.

Subculturing Procedure:

Medium Renewal: 2-3 times per week.

Subcultivation Ratio: 1:16 – 1:20 Seeding density 0.8 x10⁴cells/cm². Split subconfluent cultures (70-80%). Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 5 minutes.

Culture Conditions: Incubate the culture at 37°C with 5% CO₂.

Cryoprotectant Medium: 10% DMSO + 90% FCS

Handling Procedure for Frozen Cells: Upon receipt, frozen ampoules should be transferred directly to liquid nitrogen storage without delay, if not to be used immediately. Storage at -80°C may result in loss of viability.

Additional Information: AB12 cells are tumourigenic in syngeneic immunocompetent mice. CBA mice were also exposed to asbestos and developed tumours. When these two groups are compared, BALB/c tumours developed earlier but in fewer mice, and tumour development was evidenced by ascites but not by solid tumour formation. BALB/c cells also adapted better to in vitro conditions, with a higher proportion of cell lines relative to CBA cells.

Depositor: Richard Lake – University of Western Australia

References:

Original Reference:

Davis MR, Manning LS, Whitaker D, Garlepp MJ, Robinson BW (1992)

Establishment of a murine model of malignant mesothelioma. 

Int J Cancer 52: 881-886.

PubMed: 1459729

Cellosaurus: CVCL_4405