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modal number = 52; range = 44 to 59.
This is a hyperdiploid human cell line. The modal chromosome number was 52, but cells with 53 chromosome counts also occurred at a high frequency. The rate of cells with higher ploidies was 4.9%. Over 14 marker chromosomes were common to all cells, and 6 or more others were present in some cells. Among the common markers were: der(1)t(1;1)(p36;p21), i(2q), der(11)t(11;12)(q21;q13), t(6p12q), der(9)t(9;?;14)(p24;?q11), t(12q22q) and 8 or more others. All these markers were present in a single copy per cell. Structurally normal N13 and N14 were absent; and N1, N6, N9, N12, N17, F and G chromosomes were single. A single copy each of the X and Y chromosome was detected. The presence of the Y chromosome was confirmed by fluorescent microscopy.
The NCI-H441 cell line was derived by A.F. Gazdar, M. Brower and D. Carney and associates in 1982 from the pericardial fluid of a patient with papillary adenocarcinoma of the lung.
The cell line expresses mRNA and protein of the major surfactant apoprotein (SP-A).
Electron microscopy shows multilamellar bodies and cytoplasmic structures resembling clara cell granules.
The cells can be cloned in soft agar with or without serum.
The line has been used as a transfection host for expression of pulmonary surfactant protein (SP-B).
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 Growth Conditions: :If desired, the serum can be reduced to 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.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended Medium Renewal: 2 to 3 times per week
Preservation:
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Doubling Time:
58 hrs in medium with serum; 99 to 138 hrs in serum-free medium
Related Products:
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2001
recommended serum:ATCC 30-2020
References:
22250: Bepler G, et al. Expression of p64c-myc and neuroendocrine properties define three subclasses of small cell lung cancer. Oncogene 4: 45-50, 1989. PubMed: 2536917 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 22465: Broers JL, et al. Spontaneous changes in intermediate filament protein expression patterns in lung cancer cell lines. J. Cell Sci. 91: 91-108, 1988. PubMed: 2473086 22705: O'Reilly MA, et al. Differential effects of glucocorticoid on expression of surfactant proteins in a human lung adenocarcinoma cell line. Biochim. Biophys. Acta 970: 194-204, 1988. PubMed: 3382698 22998: O'Reilly MA, et al. In vitro translation, post-translational processing and secretion of pulmonary surfactant protein B precursors. Biochim. Biophys. Acta 1011: 140-148, 1989. PubMed: 2713400 23081: Gazdar AF, et al. Peripheral airway cell differentiation in human lung cancer cell lines. Cancer Res. 50: 5481-5487, 1990. PubMed: 2386953 23342: Baatz JE, et al. Utilization of modified surfactant-associated protein B for delivery of DNA to airway cells in culture. Proc. Natl. Acad. Sci. USA 91: 2547-2551, 1994. PubMed: 8146151 24389: . . Lung Cancer 4: 155-161, 1988. 32583: Tamura T, Stadtman TC. A new selenoprotein from human lung adenocarcinoma cells: purification, properties, and thioredoxin reductase activity. Proc. Natl. Acad. Sci. USA 93: 1006-1011, 1996. PubMed: 8577704 32830: Yamaguchi Y, et al. Biochemical characterization and intracellular localization of the Menkes disease protein. Proc. Natl. Acad. Sci. USA 93: 14030-14035, 1996. PubMed: 8943055
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