ATCC Number：CRL-9609?
相關(guān)疾?。赫?br />
細(xì)胞形態(tài)：上皮樣
細(xì)胞類型：其他細(xì)胞類型
器官來源：肺
是否是腫瘤細(xì)胞：0
物種來源：人
數(shù)量：大量
運(yùn)輸方式：凍存運(yùn)輸
組織來源：bronchus
生長(zhǎng)狀態(tài)：貼壁生長(zhǎng)
規(guī)格：0.1ml Designations: BEAS-2B
Depositors: ?The United States of America
Biosafety Level:2 [Cells contain polyomavirus DNA sequences ]
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Homo sapiens
Morphology:epithelial
Source: Organ: lung
Tissue: bronchus
Disease: normal
Cell Type: epithelialvirus transformed
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Applications:transfection host
Tumorigenic:No
DNA Profile (STR):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
Comments: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.
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.
The cells stain positively for keratins and SV40 T antigen.
Propagation: ATCC 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.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Growth Conditions: The flasks used should be precoated with 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 .
Subculturing: Protocol:
- Remove and discard culture medium.
- 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).
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 37C to facilitate dispersal. - Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
- Transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
- Discard supernatant and resuspend cells in fresh growth medium. Inoculate new flasks at 1500 to 3000 cells per sq. cm. 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 0.01mg/ml bovine serum albumin dissolved in BEBM medium (see reference below).
- Place culture flasks in incubators at 37C.
Interval: Subcultured before reaching confluence.
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium supplemented with 1% PVP and 7.5% DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products:0.25% (w/v) Trypsin - 0.53 mM EDTA in Hank' BSS (w/o Ca++, Mg++):ATCC 30-2101
Cell culture tested DMSO:ATCC 4-X
References: 21937: Reddel RR, et al. Immortalized human bronchial epitherial mesothelial cell lines. US Patent 4,885,238 dated Dec 5 1989
22301: 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.
30067: 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
