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當(dāng)前位置: 首頁 > ATCC代理 > NL20 B165399
NL20
NL20
規(guī)格:
價(jià)格:
編號(hào):B165399
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板


產(chǎn)品名稱
NL20
商品貨號(hào)
B165399
Organism
Homo sapiens, human
Tissue
lung/bronchus
培養(yǎng)基
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
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
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.

  1. Remove and discard culture medium.
  2. 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.
  3. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  4. Add appropriate aliquots of the cell suspension to new culture vessels.
  5. 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.

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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