A549 human lung carcinoma cells

Manufactured by American Type Culture Collection

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A549 human lung carcinoma cells are a well-established cell line derived from a human lung adenocarcinoma. They are adherent, epithelial-like cells that can be used for in vitro studies related to lung biology and disease.

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A549 cells (507 citations) A549 lung carcinoma (11 citations) Human A549 cells (5 citations) A549 human lung carcinoma (4 citations) A549 lung carcinoma cells (2 citations)

9 protocols using a549 human lung carcinoma cells

A549 Lung Cancer Cell Culture and Cytokine Assay

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A549 human lung carcinoma cells (catalog no. CCL‐185) were purchased from ATCC (Manassas, VA) and grown in DMEM (catalog no. 11995‐065) supplemented with 10% fetal bovine serum (catalog no. 03‐600‐511), 100 U/mL penicillin, and 100 U/mL streptomycin (ThermoFisher Scientific, Waltham, MA). Cells were maintained at 37°C in a 5% CO₂ controlled incubator and were passaged every 3 to 4 days at approximately 80% confluence. Resazurin sodium salt (ThermoFisher Scientific) was resuspended at 0.16 mg/mL in pH 7.4 phosphate‐buffered saline (PBS) and passed through a 0.2 μm syringe filter. Human NAMPT‐specific carboxyl‐terminal enzyme immunoassays (catalog no. EK‐003‐80) were purchased from Phoenix Pharmaceuticals (Burlingame, CA). High performance liquid chromatography purified NAMPT siRNA (sense: 5′‐CCACCCAACACAAGCAAAGUUUAUU‐3′) and scrambled control siRNA (sense: 5′‐CCACAACAACAAACGUUGAUCCAUU‐3′) were custom synthesized (ThermoFisher Scientific). Multiplex human cytokine magnetic bead panels for the detection of IL‐1α, IL‐1β, IL‐1Ra, IL‐6, and TNFα (MILLIPLEX MAP, HCYTOMAG‐60K) were purchased from EMD Millipore (Darmstadt, Germany).

Funk R., Singh R., Pramann L., Gigliotti N., Islam S., Heruth D., Ye S., Chan M., Leeder J, & Becker M. (2016). Nicotinamide Phosphoribosyltransferase Attenuates Methotrexate Response in Juvenile Idiopathic Arthritis and In Vitro. Clinical and Translational Science, 9(3), 149-157.

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A549 Cell Culture and NAMPT Knockdown

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A549 human lung carcinoma cells (catalogue no. CCL-185) were purchased from ATCC (Manassas, VA) and grown in DMEM (catalogue no. 11995-065) supplemented with 10% fetal bovine serum (catalogue no. 03-600-511), 100 U/mL penicillin, and 100 U/mL streptomycin (ThermoFisher Scientific, Waltham, MA). Cells were maintained at 37°C in a 5% CO₂ controlled incubator and were passaged every 3 to 4 days at ∼80% confluence. Resazurin sodium salt (ThermoFisher Scientific, Waltham, MA) was re-suspended at 0.16 mg/mL in pH 7.4 PBS and passed through a 0.2 μm syringe filter. Human NAMPT specific carboxyl-terminal enzyme immunoassays (catalogue no. EK-003-80) were purchased from Phoenix Pharmaceuticals (Burlingame, CA). High performance liquid chromatography purified NAMPT siRNA (sense: 5′-CCACCCAACACAAGCAAAGUUUAUU-3′) and scrambled control siRNA (sense: 5′-CCACAACAACAAACGUUGAUCCAUU-3′) were custom synthesized (ThermoFisher Scientific, Waltham, MA). Multiplex human cytokine magnetic bead panels for the detection of IL-1α, IL-1β, IL-1Ra, IL-6, and TNFα (MILLIPLEX MAP, HCYTOMAG-60K) were purchased from EMD Millipore (Darmstadt, Germany).

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Lung Carcinoma Cell Line Acquisition

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KB cells, a subline of HeLa cells, were obtained from the Institute of Development, Aging and Cancer, Tohoku University (Sendai, Japan). A549 human lung carcinoma cells were purchased from American Type Culture Collection (Rockville, MD, USA).

Sakai A., Yamashita Y., Misumi S., Kishimoto N., Onodera R., Higashi T., Arima H, & Motoyama K. (2022). Nanoparticles of folic acid‐methyl‐β‐cyclodextrin (FA‐MβCD)/adamantane‐albumin exhibit enhanced antitumor activity compared with FA‐MβCD alone. FEBS Open Bio, 13(2), 233-245.

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A549 Cell Culture Protocol

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A549 human lung carcinoma cells (American Type Culture Collection, Manassas, VA, USA) were grown in advanced Dulbecco's modified Eagle's medium (DMEM) containing high glucose, non-essential amino acids, sodium pyruvate, phenol red and no L-glutamine (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA). The advanced DMEM was further supplemented with 1% (v/v) penicillin and streptomycin solution (Invitrogen; Thermo Fisher Scientific, Inc.) and 10% (v/v) foetal bovine serum (Invitrogen; Thermo Fisher Scientific, Inc.). All cell cultures were maintained in a humidified atmosphere in an incubator at 37°C containing 5% CO₂, with media being changed every 48 h to ensure optimal growth.

Chudasama D., Burnside N., Beeson J., Karteris E., Rice A, & Anikin V. (2017). Perioperative detection of circulating tumour cells in patients with lung cancer. Oncology Letters, 14(2), 1281-1286.

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Culturing Human Cancer Cell Lines

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The human tumor cell lines (A375 human melanoma cells, Cat no. ATCC^® CRL-1619™; A549 human lung carcinoma cells, Cat no. ATCC^® CCL-185™; and MDA-MB-231 human breast carcinoma cells, Cat no. ATCC^® HTB-26™) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) via LGC Standards GmbH, Wesel Germany. The B164A5 murine melanoma cell line and cell culture media, and specific supplements were purchased from Sigma-Aldrich Co. (Ayrshire, UK). Human keratinocytes (HaCaT) were a gift from the University of Debrecen, Debrecen, Hungary. The cells were cultured in Dulbecco's modified Eagle's Medium (DMEM) containing 4.5 g/l glucose, L-glutamine and NaHCO₃, and supplemented with 100 U/ml penicillin, 100 µg/ml streptomycin and 10% fetal bovine serum (FBS). Cells were kept under standard conditions (humidified atmosphere, 5% CO₂, 37°C) and passaged every second day. Cell number was determined by using the cell counting chamber (Neubauer) in the presence of Trypan blue.

Mioc M., Soica C., Bercean V., Avram S., Balan-Porcarasu M., Coricovac D., Ghiulai R., Muntean D., Andrica F., Dehelean C., Spandidos D.A., Tsatsakis A.M, & Kurunczi L. (2017). Design, synthesis and pharmaco-toxicological assessment of 5-mercapto-1,2,4-triazole derivatives with antibacterial and antiproliferative activity. International Journal of Oncology, 50(4), 1175-1183.

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Culturing Human Cell Lines

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HEK293A human embryonic kidney cells and A549 human lung carcinoma cells were obtained from American Type Culture Collection (ATCC, Manassas, VA). Cells were cultured with Dulbecco’s-modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% Penicillin Streptomycin Glutamine (PSG).

Mendez N., Herrera V., Zhang L., Hedjran F., Feuer R., Blair S., Trogler W., Reid T, & Kummel A. (2014). Encapsulation of Adenovirus Serotype 5 in Anionic Lecithin Liposomes using a Bead-Based Immunoprecipitation Technique Enhances Transfection Efficiency. Biomaterials, 35(35), 9554-9561.

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Luciferase-Based Ad26 Neutralization Assay

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Ad26 neutralizing antibody titers in serum were assessed using a luciferase-based VNA. Briefly, an E1/E3–deleted Ad26-luciferase reporter construct was added to 96-well half-area tissue culture–treated plates (Greiner) at a multiplicity of infection of 1,000, together with twofold serial dilutions of individual heat-inactivated serum samples starting at a 1:16 dilution in a total volume of 25 µl. After 1 h, A549 human lung carcinoma cells (catalog number ATCC CCL-185, obtained from the American Type Culture Collection) were added at a density of 1 × 10⁴ cells/well. After incubation for 20 h at 37°C and 10% CO₂, luciferase activity was measured using the Neo-Lite Luciferase Assay System (Perkin Elmer), and an EnVision multimode plate reader (Perkin Elmer). 90% neutralization titers (IC₉₀) were defined as the maximum serum dilution that neutralized 90% of luciferase activity. Each serum sample was analyzed in duplicate.

Solforosi L., Kuipers H., Jongeneelen M., Rosendahl Huber S.K., van der Lubbe J.E., Dekking L., Czapska-Casey D.N., Izquierdo Gil A., Baert M.R., Drijver J., Vaneman J., van Huizen E., Choi Y., Vreugdenhil J., Kroos S., de Wilde A.H., Kourkouta E., Custers J., van der Vlugt R., Veldman D., Huizingh J., Kaszas K., Dalebout T.J., Myeni S.K., Kikkert M., Snijder E.J., Barouch D.H., Böszörményi K.P., Stammes M.A., Kondova I., Verschoor E.J., Verstrepen B.E., Koopman G., Mooij P., Bogers W.M., van Heerden M., Muchene L., Tolboom J.T., Roozendaal R., Brandenburg B., Schuitemaker H., Wegmann F, & Zahn R.C. (2021). Immunogenicity and efficacy of one and two doses of Ad26.COV2.S COVID vaccine in adult and aged NHP. The Journal of Experimental Medicine, 218(7), e20202756.

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Culturing HEK293 and A549 Cell Lines

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Human Embryonic Kidney (HEK) 293 cell line and human A549 lung carcinoma cells, obtained from the American Type Culture Collection (ATCC), were grown in Dulbecco’s modified Eagle’s medium (DMEM), (Bio-Whittaker, Cambrex Bio Science, Verviers, Belgium), supplemented with 10% fetal bovine serum (FBS), (Biological Industries, Kibbutz Beit Haemek, Israel), 1mM glutamine and antibiotics (penicillin 100 U/mL, streptomycin 100 µg/mL). All cells were maintained at 37 °C in a 5% CO₂ atmosphere.

Marsili G., Acchioni C., Remoli A.L., Amatore D., Sgarbanti R., De Angelis M., Orsatti R., Acchioni M., Astolfi A., Iraci N., Puzelli S., Facchini M., Perrotti E., Cecchetti V., Sabatini S., Superti F., Agamennone M., Barreca M.L., Hiscott J., Nencioni L, & Sgarbanti M. (2023). Identification of Anti-Influenza A Compounds Inhibiting the Viral Non-Structural Protein 1 (NS1) Using a Type I Interferon-Driven Screening Strategy. International Journal of Molecular Sciences, 24(13), 10495.

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Culturing A549 and IMR-90 Lung Cells

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Human A549 lung carcinoma cells and IMR-90 lung fibroblasts were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). These two cell lines were cultured in Dulbecco’s modified Eagle medium (DMEM, Gibco Laboratories, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS, Biological Industries, Kibbutz Beit Haemek, Israel). All cells were grown at 37 °C with 5% CO₂.

Wang W.H., Hsu C.L., Huang H.C, & Juan H.F. (2020). Quantitative Phosphoproteomics Reveals Cell Alignment and Mitochondrial Length Change under Cyclic Stretching in Lung Cells. International Journal of Molecular Sciences, 21(11), 4074.

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