| 產品名稱 |
HBE4-E6/E7-C1 [NBE4-E6/E7-C1] |
| 商品貨號 |
B164549 |
| Organism |
Homo sapiens, human |
| Tissue |
lung; bronchus; epithelial |
| Cell Type |
epithelial; human papillomavirus 16 (HPV-16) E6/E7 transforme |
| 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. |
| Age |
60 years |
| Gender |
male |
| Ethnicity |
Caucasian |
| Storage Conditions |
liquid nitrogen vapor phase |
| Karyotype |
45, X, -Y, dup (5), -8, +9, -14, -15, -20, -21, -22, +mar1, +mar2, +der(8q;13q) |
| Derivation |
The HBE4-E6/E7-C1 (formerly NBE4-E6/E7-C1) cell line was cloned from passage 22 of HBE4-E6/E7 (see ATCC CRL-2078) by limiting dilution. |
| Clinical Data |
male Caucasian 60 years |
| Comments |
The HBE4-E6/E7-C1 (formerly NBE4-E6/E7-C1) cell line was cloned from passage 22 of HBE4-E6/E7 (see ATCC CRL-2078) by limiting dilution. The cloned line is more sensitive than the parental line to induction of terminal differentiation by phorbol esters. The cells are non-viable in DMSO and should be frozen in culture medium with 10% glycerol. |
| 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 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution containing 0.5% 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 containing 0.1% soybean trypsin inhibitor and 0.1% bovine serum albumin and aspirate cells by gently pipetting.
- To remove trypsin-EDTA solution, 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 culture medium. Add appropriate aliquots of cell suspension to new culture vessels.
- Place culture vessels in incubators at 37°C.
Subcultivation Ratio: 1:2 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, 3rd edition, published by Alan R. Liss, N.Y., 1994. |
| Cryopreservation |
Complete growth medium described above supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X. |
| Culture Conditions |
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5% |
| Population Doubling Time |
24 hrs |
| Name of Depositor |
J Viallet |
| Deposited As |
Homo sapiens |
| Passage History |
The HBE4-E6/E7-C1 (formerly NBE4-E6/E7-C1) cell line was cloned from passage 22 of HBE4-E6/E7 (see ATCC CRL-2078) by limiting dilution. |
| 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
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.
|
