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HBE4-E6/E7 [NBE4-E6/E7] (ATCC® CRL-2078™)

Organism: Homo sapiens, human  /  Cell Type: epithelial; human papillomavirus 16 (HPV-16) E6/E7 transformed  /  Tissue: lung; bronchus  /  Disease: normal

Organism Homo sapiens, human
Tissue lung; bronchus
Cell Type epithelial; human papillomavirus 16 (HPV-16) E6/E7 transformed
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2  [Cells contain human papilloma viral DNA sequences]

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease normal
Age 60 years
Gender male
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Karyotype 45, X, -Y, dup (5), -8, +9, -20, -22, +mar1, +mar2
Derivation
The HBE4-E6/E7 (formerly NBE4-E6/E7) cell line was derived from normal bronchial epithelium taken from a man undergoing a left upper lobectomy for a poorly differentiated (T2 N0) adenocarcinoma.
Cells from the primary explant in their first passage were transfected with the p1321 plasmid encoding the E6 and E7 genes of Human Papilloma virus type 16 (HPV 16) under the control of the human beta actin promotor.
Clinical Data
60 years
Caucasian
male
Tumorigenic No
Effects
No, not tumorigenic in nude mice
Comments
These sequences are known to bind to and inactivate endogenous p53 and Rb proteins respectively.
Southern blot analysis shows that stable integration of the transfected genes occurred.
The cell line resembles morphologically the basal cells of the normal human bronchial epithelium, and is not tumorigenic in athymic nude mice.
The cells are able to form tubules when grown in a basement membrane like matrix, and they retain the ability to undergo terminal squamous differentiation in response to phorbol esters or upon reaching confluence.
Cells are non-viable in DMSO and should be frozen in culture Medium with 10% glycerol.
Original authentication testing at ATCC by isoenzymology indicated that this was a human cell line. Recent additional speciation using a mitochondrial cytochrome c oxidase I (CO1) assay has shown that this cell line is cross-contaminated with bovine cells. Additional more sensitive PCR assays of the original token lot have also indicated a low level of bovine cross-contamination in that material as well.
Complete Growth Medium The base medium for this cell line is provided by Invitrogen (GIBCO) as part of a kit: Keratinocyte Serum Free Medium (K-SFM), Kit Catalog Number 17005-042. This kit is supplied with two additives required to grow this cell line (bovine pituitary extract (BPE) and human recombinant epidermal growth factor (EGF). To make the complete growth medium, you will need to add the following components to the base medium:
  • 0.05 mg/ml BPE - provided with the K-SFM kit
  • 5 ng/ml EGF - provided with the K-SFM kit
  • 10 ng/ml cholera toxin - not provided with kit
NOTE: Do not filter complete medium
Subculturing
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution containing 0.5% polyvinylpyrrolidone (PVP).
  3. 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.
  4. Add 2.0 to 3.0 mL of complete growth medium containing 0.1% soybean trypsin inhibitor and 0.1% bovine serum albumin and aspirate cells by gently pipetting.Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
  5. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.
Subculture before the cells become confluent.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: Every 2 to 3 days
Cryopreservation
culture medium, 90%; glycerol, 10%
Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
STR Profile
Amelogenin: X,Y
CSF1PO: 11,12
D13S317: 11
D16S539: 11,13
D5S818: 10,11
D7S820: 10,11
THO1: 6,8
TPOX: 8
vWA: 14,17
Population Doubling Time 24 hrs
Name of Depositor J Viallet
Deposited As Homo sapiens
Passage History
Cells from the primary explant in their first passage were transfected with the p1321 plasmid encoding the E6 and E7 genes of Human Papilloma virus type 16 (HPV 16) under the control of the human beta actin promotor.
References

Viallet J, et al. Characterization of human bronchial epithelial cells immortalized by the E6 and E7 genes of human papillomavirus type 16. Exp. Cell Res. 212: 36-41, 1994. PubMed: 8174640

Tsao MS, et al. Autocrine growth loop of the epidermal growth factor receptor in normal and immortalized human bronchial epithelial cells. Exp. Cell Res. 223: 268-273, 1996. PubMed: 8601403

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., 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
Product Sheet
Product Sheet
Certificate of Analysis
Certificate of Analysis
SDS
SDS
References

Viallet J, et al. Characterization of human bronchial epithelial cells immortalized by the E6 and E7 genes of human papillomavirus type 16. Exp. Cell Res. 212: 36-41, 1994. PubMed: 8174640

Tsao MS, et al. Autocrine growth loop of the epidermal growth factor receptor in normal and immortalized human bronchial epithelial cells. Exp. Cell Res. 223: 268-273, 1996. PubMed: 8601403

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., 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.