BEAS-2B (ATCC® CRL-9609)

Organism: Homo sapiens, human  /  Cell Type: epithelial (virus transformed)  /  Tissue: lung/bronchus  /  Disease: normal

Organism Homo sapiens, human
Tissue lung/bronchus
Cell Type epithelial (virus transformed)
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain polyomavirus DNA sequences]
Disease normal
Applications

This cell line is a suitable transfection host.

The cells retain the ability to undergo squamous differentiation in response to serum, and can be used to screen chemical and biological agents for ability to induce or affect differentiation and/or carcinogenesis.

Storage Conditions liquid nitrogen vapor phase
Images
Derivation
Epithelial cells were isolated from normal human bronchial epithelium obtained from autopsy of non-cancerous individuals.

The cells were infected with an adenovirus 12-SV40 virus hybrid (Ad12SV40) and cloned.
Clinical Data
Epithelial cells were isolated from normal human bronchial epithelium obtained from autopsy of healthy individuals.
Tumorigenic The cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
Effects
Yes, the cells did form colonies in semisolid medium
No, the cells were not tumorigenic in immunosuppressed mice
Comments
The cells stain positively for keratins and SV40 T antigen.


Complete Growth Medium The base medium for this cell line (BEBM) along with all the additives can be obtained from Lonza/Clonetics Corporation as a kit: BEGM, Kit Catalog No. CC-3170. ATCC does not use the GA-1000 (gentamycin-amphotericin B mix) provided with the BEGM kit. Note: Do not filter complete medium.
Subculturing

These cells should be subcultured before reaching confluence since confluent cultures rapidly undergo squamous terminal differentiation. Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. Add 2.0 to 3.0 mL of 0.25% Trypsin - 0.53mM EDTA solution containing 0.5% polyvinylpyrrolidone (PVP) to flask and observe cells under an inverted microscope until cell layer is dispersed (usually with 5 to 10 minutes).
  3. Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.  Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4.  Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. 
  5. Transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes
  6. Discard supernatant and resuspend cells in fresh growth medium. Inoculate new flasks at 1500 to 3000 cells per cm2.  The culture flasks used should be pre-coated with a mixture of 0.01mg/ml fibronectin, 0.03 mg/ml bovine collagen type I and 0.01 mg/mL bovine serum albumin dissolved in BEBM.  
  7. Place culture flasks in incubators at 37°C.

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Interval: Subcultured before reaching confluence.
Medium Renewal: Every 2 to 3 days

Flask Coating
 
  1. Prepare a mixture of 0.01 mg/mL fibronectin, 0.03 mg/mL bovine collagen type I and 0.01 mg/mL bovine serum albumin (BSA) dissolved in culture medium. Store pre-prepared Coating Solution at 4°C in cold room for up to 3 months. 
  2. For a growth area of 75 cm2, add 4.5 mL of the fibronectin/collagen/BSA solution and rock gently to coat the entire surface. 
  3. Incubate the freshly coated vessel(s) in a 37°C incubator overnight (it is preferable to use tissue culture vessels with tightened, plug-seal caps to prevent evaporation during the coating process). 
  4. Store coated flasks with solution at room temperature, light protected, up to 1 month. Suction off solution before plating cells.
Cryopreservation
Freeze medium: Complete growth medium supplemented with 1% PVP and 7.5% DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Growth Conditions: The culture flasks used should be pre-coated with a mixture of 0.01 mg/mL fibronectin, 0.03 mg/mL bovine collagen type I and 0.01 mg/mL bovine serum albumin dissolved in BEBM medium (see references: U.S. Pat. 4,885,238 and Lechner, J.F. and LaVeck, M.A. A serum-free method for culturing normal bronchial epithelial cells at clonal density. J. Tissue Culture Methods 9: 4348, 1985)
STR Profile
Amelogenin: XY
CSF1PO: 9, 12
D13S317: 13
D16S539: 12
D5S818: 12,13
D7S820: 10, 13
THO1: 7, 9.3
TPOX: 6, 11
vWA: 17, 18
Name of Depositor The United States of America
References

Reddel RR, et al. Immortalized human bronchial epitherial mesothelial cell lines. US Patent 4,885,238 dated Dec 5 1989

Lechner JF, LaVeck MA. A serum-free method for culturing normal human bronchial epithelial cells at clonal density. J. Tissue Culture Methods 9: 43-48, 1985.

Sakamoto O, et al. Role of macrophage-stimulating protein and its receptor, RON tyrosine kinase, in ciliary motility. J. Clin. Invest. 99: 701-709, 1997. PubMed: 9045873

Hay RJ, Caputo JL, Macy, ML, Eds. (1992) ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo JL. Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Basic Documentation
Other Documentation
References

Reddel RR, et al. Immortalized human bronchial epitherial mesothelial cell lines. US Patent 4,885,238 dated Dec 5 1989

Lechner JF, LaVeck MA. A serum-free method for culturing normal human bronchial epithelial cells at clonal density. J. Tissue Culture Methods 9: 43-48, 1985.

Sakamoto O, et al. Role of macrophage-stimulating protein and its receptor, RON tyrosine kinase, in ciliary motility. J. Clin. Invest. 99: 701-709, 1997. PubMed: 9045873

Hay RJ, Caputo JL, Macy, ML, Eds. (1992) ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo JL. Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.