HBE135-E6E7 (ATCC® CRL-2741)

Organism: Homo sapiens, human  /  Cell Type: epithelial HPV-16 E6/E7 transformed  /  Tissue: lung/bronchus  /  Disease: lung cancer

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Organism Homo sapiens, human
Tissue lung/bronchus
Cell Type epithelial HPV-16 E6/E7 transformed
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain HPV-16 E6/E7 viral DNA sequences]
Disease lung cancer
Age 54 years
Gender male
Applications
This cell line may be used as a reference in cDNA microarray studies to demonstrate lung cancer profiles correlating with clinical outcome or biology of tumors. RefWigle DA, et al. Molecular profiling of non-small cell lung cancer and correlation with disease-free survival. Cancer Res. 62: 3005-3008, 2002. PubMed: 12036904 
Storage Conditions liquid nitrogen vapor phase
Derivation
The HBE135-E6E7 cell line was derived from normal bronchial epithelium taken from a man undergoing lobectomy for squamous cell carcinoma. Cells from the primary explant in their first passage were infected with the recombinant retrovirus LXSN16E6E7 containing the human papilloma virus (HPV) E6E7 gene. Cells were selected in the presence of 0.4 mg/mL G418. RefTsao 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
Clinical Data
54 years
male
Comments

Northern blot hybridization shows that immortalized HBE cells express high levels of mRNA for epidermal growth factor receptor (EGFR), transforming growth factor-alpha (TGF-alpha), and amphiregulin (AR), but not epidermal growth factor (EGF). RefTsao 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

 

 

Complete Growth Medium Keratinocyte-Serum Free medium with 5 ng/ml human recombinant EGF (do not filter) and 0.05 mg/ml bovine pituitary extract (Invitrogen, formerly GIBCO-BRL, Cat. No. 17005-042) and supplemented with 0.005 mg/ml insulin and 500 ng/ml hydrocortisone.
Subculturing 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. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA 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 6.0 to 8.0 mL of complete growth medium and aspirate cells by pipetting gently.
  5. To remove trypsin-EDTA solution, 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 serum-free growth medium.  Add appropriate aliquots of cell suspension to new culture vessels.
  7. Place culture vessels in incubators at 37°C.

Subcultivation Ratio: 1:3 to 1:4
Medium Renewal: Every 2 to 3 days.  
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, 3th edition, published by Alan R. Liss, N.Y., 1994.
Cryopreservation
Freeze medium: Complete growth medium supplemented with 10% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Population Doubling Time 24 to 30
Name of Depositor M Tsao
Passage History
Cells from the primary explant in their first passage were infected with the recombinant retrovirus LXSN16E6E7 containing the human papilloma virus (HPV) E6E7 gene.
Year of Origin 1994
References

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

The cell line was established from normal bronchus tissue taken at lobectomy for squamous cell carcinoma. The tissue used was at least 3 cm from the edge of the tumor.

Wigle DA, et al. Molecular profiling of non-small cell lung cancer and correlation with disease-free survival. Cancer Res. 62: 3005-3008, 2002. PubMed: 12036904

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

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

The cell line was established from normal bronchus tissue taken at lobectomy for squamous cell carcinoma. The tissue used was at least 3 cm from the edge of the tumor.

Wigle DA, et al. Molecular profiling of non-small cell lung cancer and correlation with disease-free survival. Cancer Res. 62: 3005-3008, 2002. PubMed: 12036904