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Isolation: Isolation date: 1982
Tumorigenic: Yes
DNA Profile (STR): Amelogenin: X,Y
CSF1PO: 11,12
D13S317: 13
D16S539: 9
D5S818: 9,10
D7S820: 9,12
THO1: 9.3
TPOX: 8
vWA: 17
Cytogenetic Analysis: modal numbr = 57; range = 53 to 65.This is a hypotriploid human cell line. The modal chromosome number is 57 although cells with 58 chromosomes occurred with a comparable frequency. The frequency of higher ploidies was 1.7%. Seven marker chromosomes, der(9)t(1;9)(q21;p24), der(9)t(7;9)(p11;p22), t(10q14q), der(16)t(7;16)(q11.23;q22), a small ring (about 1/2 the size of a G chromosome) and two others, were common to all cells. Three other markers were found in some cells only. The markers, t(7;9) and t(7;16) were mostly paired. Normal N9 was absent, and N7 and N16 had only a single copy per cell. Two copies each of the X and the Y were present in all cells.
Isoenzymes: AK-1, 1
ES-D, 1
G6PD, B
GLO-I, 1-2
Me-2, 1
PGM1, 1
PGM3, 1
Gender: male
Comments: The NCI-H460 cell line was derived by A.F. Gazdar and associates in 1982 from the pleural fluid of a patient with large cell cancer of the lung.
The cells express easily detectable p53 mRNA at levels comparable to normal lung tissue, and exhibit no gross structural DNA abnormalities.
The cells stain positively for keratin and vimentin but are negative for neurofilament triplet protein.
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Temperature: 37.0°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Subculturing: Protocol:
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
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 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.
Subc*tion Ratio: A subc*tion ratio of 1:3 to 1:8 is recommended
Medium Renewal: Twice per week
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Doubling Time: 23 hrs in medium with serum; 42 to 60 hrs in serum
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2001
recommended serum:ATCC 30-2020
References: 1605: Banks-Schlegel SP, et al. Intermediate filament and cross-linked envelope expression in human lung tumor cell lines. Cancer Res. 45: 1187-1197, 1985. PubMed: 2578876
1806: Takahashi T, et al. p53: A frequent target for genetic abnormalities in lung cancer. Science 246: 491-494, 1989. PubMed: 2554494
22434: Brower M, et al. Growth of cell lines and clinical specimens of human non-small cell lung cancer in a serum-free defined medium. Cancer Res. 46: 798-806, 1986. PubMed: 3940644
32488: Geiger T, et al. Antitumor activity of a PKC-alpha antisense oligonucleotide in combination with standard chemotherapeutic agents against various human tumors transplanted into nude mice. Anticancer Drug Des. 13: 35-45, 1998. PubMed: 9474241
