Description: ONE58 is a human mesothelioma cell line derived from a male patient. The patient had known exposure to crocidolite asbestos. The culture was developed from a pleural effusion sample. Malignant mesothelioma was diagnosed through analysis of pleural effusion fluid.

Organism: Human (Homo sapiens)

Strain: N/A

Tissue: Pleural Fluid

Growth Properties: Adherent

Morphology: Epithelial-like: cells are spindle-shaped with few vacuoles.

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: 1:4-1:8. Seeding density 1.2 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: ONE58 cells express cytokeratin and epithelial membrane antigen (EMA) but not mucin. ONE58 carries point mutations in topoisomerase II-beta. These mutations are – L532L, S563P.

Depositor: Richard Lake – University of Western Australia, Australia.

Reference:

Manning LS, Whitaker D, Murch AR, Garlepp MJ, Davis MR, Musk AW, Robinson BW (1991) Int J Cancer Jan 21;47(2):285-90

PubMed 1703129

Cellosaurus: CVCL_2671