Description: BEAS-2B cells were derived from normal bronchial epithelium obtained from autopsy of non-cancerous individuals. Cells were infected with a replication-defective SV40/adenovirus 12 hybrid and cloned. Squamous differentiation can be observed in response to serum. This ability can be used for screening chemical and biological agents inducing or affecting differentiation and/or carcinogenesis. The cell line has been applied for studies of pneumococcal infection mechanisms. BEAS-2B was described to express keratins and SV40 T antigen. Subculturing the cells before confluency is necessary as confluent cultures rapidly undergo squamous terminal differentiation. This material is cited in a U.S and / or other Patent (U.S. Pat. 4,885,238) and may not be used to infringe the patent claims.

Also Known As:

Species: Human

Tissue: Lung

Growth Properties: Adherent

Morphology: Epithelial-like

Growth Medium: BEGM, also known as LHC-9 with modification (available from Clonetics Corporation, CC3171 (BEBM) plus additives CC4175 or BEGM Bullet Kit, CC3170); Note: Additives supplied contain gentamycin which has been omitted for culture at ECACC; media is serum-free.

Subculturing Procedure: A centrifugation step to remove the cryoprotectant is essential. Rapidly thaw the frozen ampoule in a water bath at 37°C for 1-2 minutes. Transfer the contents to a centrifuge tube and slowly add 5-10ml of pre-warmed growth media. Remove a sample for counting. Centrifuge at 100-150g for 2-3 minutes to pellet cells and seed at10, 000 cells per cm². Check cultures daily  and passage sub-confluent cultures (maximum 70% confluence )using 0.25% trypsin or trypsin/EDTA. Incubate at room temperature for 5-10 min until cells detach. Add fresh medium and disperse cells, centrifuge and resuspend pellet in medium. Seed into new flasks between 3000 and 10, 000 cells per cm². Flasks have to be precoated with a mixture of 0.01 mg/ml fibronectin, 0.03 mg/ml collagen and 0.01 mg/ml bovine serum albumin dissolved in BEGM. Add mixture at ratio of 0.2 ml per cm² surface area. Incubate at 37°C for at least 6 hours 5% CO₂. Remove excess fluid and allow flasks to dry by incubating at 37°C overnight, leaving the caps loose. Prior to addition of cells wash flask three times with PBS. Note: BEGM medium is almost serum-free, therefore trypsin inhibitor is essential. Add an equal or greater volume of trypsin inhibitor as trypsin used, centrifuge and reseed cells in collagen coated flasks using culture medium.

When freezing cells down use 50:50 fresh culture medium:conditioned medium plus 10% DMSO

Release Conditions: No

Depositor:

Originator: No

References: J. Tissue Culture Methods 1985; 9:43 Infect Immun 1998; 66:820 J In Vitro Toxicology 1997; 10:93 J In Vitro Toxicology 1997; 10:85

Hyperlink to ECACC Cell Line Data Sheet: 95102433