HTB-177 NCI-H460 人大細(xì)胞肺癌細(xì)胞

ATCC® Number:  HTB-177™

Designations:  NCI-H460 [H460]

Depositors:   AF Gazdar, JD Minna

Biosafety Level: 1

Shipped:  frozen

Medium & Serum:  See Propagation

Growth Properties: adherent

Organism: Homo sapiens (human)

Morphology: epithelial

Source: Organ: lung

Disease: carcinoma; large cell lung cancer

Derived from metastatic site: pleural effusion

Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimay responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.

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

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