Cell Line Description: AC29 is a mouse mesothelioma cell line. Mice (CBA strain, female, 6-8 weeks old) were exposed to crocidolite asbestos through intraperitoneal injection, resulting in tumour development (both ascites and solid tumours). Cultures were established from malignant mesothelial cells obtained from ascites fluid.

Organism: Mouse (Mus musculus)

Strain: CBA

Tissue: Mesothelium malignant mesothelial cells were obtained from ascites fluid

Growth Properties: Adherent

Morphology: Epithelial-like

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.

Subculturing Procedure:

Medium Renewal: 2-3 times per week.

Subcultivation Ratio: 1:8 – 1:16. Seeding density 0.6 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: Cells are tumourigenic in syngeneic immunocompetent mice. CBA cells did not adapt as well to in vitro conditions with a lower proportion of cell lines relative to BALB/c cells. AC29 cells express WT1 and have been used in the study of Wilms’ tumour.

Depositor: Richard Lake – University of Western Australia

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_4407