產(chǎn)品名稱 |
NL20 |
商品貨號 |
B165399 |
Organism |
Homo sapiens, human |
Tissue |
lung/bronchus |
Cell Type |
Epithelial |
Product Format |
frozen |
Morphology |
epithelial |
Culture Properties |
adherent |
Biosafety Level |
2 [Cells contain SV40 viral 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. |
Disease |
Accident victim |
Age |
20 years |
Gender |
female |
Ethnicity |
Caucasian, White |
Applications |
The non-tumorigenic NL20 cell line and the tumorigenic NL20-TA cell line form a pair of immortal cell lines that can be used to study tumor progression. |
Storage Conditions |
liquid nitrogen vapor phase |
Karyotype |
near diploid |
Images |
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Derivation |
NL20 (ATCC CRL-2503) is an immortalized, nontumorigenic human bronchial epithelial cell line derived from normal bronchus taken from an accident victim at autopsy.
The cell line was established by transfection with the origin of replication-defective SV40 large T plasmid, p129.
After three months the tumor was removed and placed in culture. At passage 3, these cells were re-injected into nude mice.
NL20 cells at passage 183 were inoculated into nude mice and a small slowly growing subcutaneous tumor developed from a minor clone in this otherwise stable cell line. |
Clinical Data |
20 years Caucasian, White female |
Tumorigenic |
No |
Effects |
No, The cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium. |
Comments |
NL20 cells at passage 183 were inoculated into nude mice and a small slowly growing subcutaneous tumor developed from a minor clone in this otherwise stable cell line. After three months the tumor was removed and placed in culture. At passage 3, these cells were re-injected into nude mice. One of the resulting tumors was dissociated, placed in culture and designated NL20-TA. This cell line (ATCC CRL-2504) remains tumorigenic up to at least passage 250. |
Complete Growth Medium |
Ham's F12 medium with 1.5 g/L sodium bicarbonate, 2.7 g/L glucose, 2.0 mM L-glutamine, 0.1 mM nonessential amino acids, 0.005 mg/ml insulin, 10 ng/ml epidermal growth factor, 0.001 mg/ml transferrin, 500 ng/ml hydrocortisone and 4% fetal bovine serum
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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.
- Add 10-15 mL dissociation solution (0.02% EDTA and 5% dialyzed fetal bovine serum in Ca++/Mg++ free Hanks' BSS) and allow the flask to sit at 37°C for 12 minutes or until the cells detach.
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.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C
Subculture Ratio: 1:12 to 1:20
Medium Renewal: Every 2 to 3 days.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.
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Cryopreservation |
Culture medium 95%; DMSO, 5% |
Culture Conditions |
Temperature: 37.0°C |
Name of Depositor |
JH Schiller |
Deposited As |
human |
Passage History |
One of the resulting tumors was dissociated, placed in culture and designated NL20-TA. This cell line (ATCC CRL-2504) remains tumorigenic up to at least passage 250. |
References |
Schiller JH, et al. Phenotypic, molecular and genetic characterization of transformed human bronchial epithelial cell strains. Int. J. Oncol. 4: 461-470, 1994.
Schiller JH, Bittner G. Loss of the tumorigenic phenotype with in vitro, but not in vivo, passaging of a novel series of human bronchial epithelial cell lines: possible role of an alpha 5/beta 1-integrin-fibronectin interaction. Cancer Res. : 6215-6221, 1995. PubMed: 8521416
Wittenkeller JL, et al. Comparison of spontaneous and induced mutation rates in an
immortalized human bronchial epithelial cell line and its tumorigenic derivative. Oncology 54: 335-341, 1997. PubMed: 9216860
Schiller JH, et al. Karyotypic changes associated with spontaneous acquisition and loss of tumorigenicity in a human transformed bronchial epithelial cell line: evidence for in vivo selection of transformed clones. In Vitro Cell. Dev. Biol. Anim. 34: 283-289, 1998. PubMed: 9590501
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