BBM (ATCC® CRL-9482)

Organism: Homo sapiens, human  /  Cell Type: epithelialvirus transformed  /  Disease: Carcinogen

Organism Homo sapiens, human
Cell Type epithelialvirus transformed
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 Cells contain polyomavirus DNA sequences
Disease Carcinogen
Applications
Tranformants were selected in medium containing G418. The cells can be used to screen chemical and biological agents for activity as growth factor, carcinogens, mutagens, etc.
This line was derived from BEAS-2B cells (see ATCC CRL-9609) by transfection with the B-myc/pSV2neo plasmid (constructed by ligating a BamH1/EcoR1 fragment of the c-myc gene from CA46 cells to a BamH1/EcoR1 fragment of the pSV2neo plasmid).
Storage Conditions liquid nitrogen vapor phase
Derivation
This line was derived from BEAS-2B cells (see ATCC CRL-9609) by transfection with the B-myc/pSV2neo plasmid (constructed by ligating a BamH1/EcoR1 fragment of the c-myc gene from CA46 cells to a BamH1/EcoR1 fragment of the pSV2neo plasmid). Tranformants were selected in medium containing G418. The cells can be used to screen chemical and biological agents for activity as growth factor, carcinogens, mutagens, etc. The cells are reported to stain positively for keratins and SV40 T antigen.
Tumorigenic No,
Effects
Yes, did form colonies in semisolid medium.
No, the cells were not tumorigenic in immunosuppressed mice,
Comments
This line was derived from BEAS-2B cells (see ATCC CRL-9609) by transfection with the B-myc/pSV2neo plasmid (constructed by ligating a BamH1/EcoR1 fragment of the c-myc gene from CA46 cells to a BamH1/EcoR1 fragment of the pSV2neo plasmid). Tranformants were selected in medium containing G418. The cells can be used to screen chemical and biological agents for activity as growth factor, carcinogens, mutagens, etc. The cells are reported to 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
Protocol:
  • Remove and discard culture medium.
  • Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution containing 0.5%(w/v) polyvinylpyrrolidone (PVP).
  • Add 2.0 to 3.0 ml of Trypsin-EDTA-PVP solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    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.
  • Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
  • To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
  • Incubate cultures at 37°C.
Subcultivation Ratio: Inoculate new flasks at 1500 to 3000 cells per sq. cm.
Medium Renewal: Two to three times weekly
Cryopreservation
Freeze medium: Leibovitz's L-15 medium with 2 mM L-glutamine supplemented with 10 mM HEPES, 1% PVP, 10% fetal bovine serum and 7.5% DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Growth Conditions: The flasks used should be precoated 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 reference below) (see reference below).
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.

Basic 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.