年限：54 years adult
細(xì)胞形態(tài)：上皮樣
生長狀態(tài)：貼壁生長
器官來源：肺
運(yùn)輸方式：凍存運(yùn)輸
組織來源：bronchus
細(xì)胞類型：其他細(xì)胞類型
是否是腫瘤細(xì)胞：0
物種來源：人
ATCC Number：CRL-2741?
數(shù)量：大量
規(guī)格：48T Designations: HBE135-E6E7
Depositors: ?M Tsao
Biosafety Level:2 [Cells contain HPV-16 E6/E7 viral DNA sequences ]
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Homo sapiens deposited as human
Morphology:epithelial
Source: Organ: lung
Tissue: bronchus
Cell Type: epithelialHPV-16 E6/E7 transformed
Permits/Forms:In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Isolation: Isolation date: 1994
Age: 54 years adult
Gender: male
Comments:The HBE135-E6E7 cell line was derived from normal bronchial epithelium taken from a man undergoing lobectomy for squamous cell carcinoma [PubMed: 8601403]. 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 [PubMed: 8601403]. 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) [PubMed: 8601403]. 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 [PubMed: 12036904].
Propagation: ATCC 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.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol:
- Remove and discard culture medium.
- 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.
- 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. - 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: 1:3 to 1:4
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium supplemented with 10% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Doubling Time: 24 to 30
References: 57773: 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
89587: 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
