| 產(chǎn)品名稱 |
RL-65 |
| 商品貨號(hào) |
B165607 |
| Organism |
Rattus norvegicus, rat |
| Tissue |
lung |
| Cell Type |
Squamous Epithelium,Epithelial |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
1
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 |
5 days |
| Strain |
Sprague-Dawley |
| Storage Conditions |
liquid nitrogen vapor phase |
| Disclosure |
This material is cited in a US or other Patent and may not be used to
infringe the claims. Depending on the wishes of the Depositor, ATCC may be
required to inform the Patent Depositor of the party to which the material was
furnished. This material may not have been produced or characterized by ATCC. |
| Genes Expressed |
cytoskeletal proteins (alpha keratin, actin, desmin, vimentin, tubulin); fibronectin; laminin |
| Cellular Products |
cytoskeletal proteins (alpha keratin, actin, desmin, vimentin, tubulin); fibronectin; laminin |
| Tumorigenic |
No |
| Effects |
No, in immunosuppressed mice Yes, in semisolid medium. |
| Comments |
The cells are maintained in a serum free medium. If grown in media containing serum, the properties of the cells will change.
The cells exhibit different characteristic when grown with or without retinoic acid.
In the presence of retinoic acid (50 nM), the cells resemble low non-keratinized or squamous epithelium with densely packed colonies. In the absence of retinoic acid, the cells form a keratinized epithelium.
Long term cultures in the absence of retinoic acid form a stratified highly keratinized epithelium with large networks of epithelial filaments.
|
| Complete Growth Medium |
A 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium with 2.5 mM L-glutamine adjusted to contain 1.2 g/L sodium bicarbonate and supplemented with 0.005 mg/ml insulin, 0.01 mg/ml human transferrin, 0.1 mM ethanolamine, 0.1 mM phosphoethanolamine, 25 nM selenium, 500 nM hydrocortisone, 0.005 mM forskolin, and bovine pituitary extract (0.15 mg protein per ml).
|
| 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.
- 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 culture medium and aspirate cells by gently pipetting.
- Transfer the cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 7 minutes.
- Resuspend the cell pellet with the recommended complete growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:10 to 1:20 is recommended
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 culture medium 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X. |
| Culture Conditions |
Temperature: 37°C Atmosphere: Air, 95%; Carbon dioxide (CO2), 5% |
| Name of Depositor |
Genentech, Inc. |
| Deposited As |
Rattus sp. |
| U.S. Patent Number |
|
| References |
Mather JP, Roberts PE. Method of isolating lung cell line. US Patent 5,364,785 dated Nov 15 1994
Roberts PE, et al. A novel epithelial cell from neonatal rat lung: isolation and differentiated phenotype. Am. J. Physiol. 259: L415-L425, 1990. PubMed: 2260675
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.
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