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Atcc ccl 185tm

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ATCC® CCL-185TM is a cell line derived from human lung carcinoma. It is a widely used in vitro model for cancer research.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Penicillin streptomycin, by Merck Group (2 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Crl 5803, by American Type Culture Collection (1 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Mrc 5, by American Type Culture Collection (1 mentions) Mda mb 231, by American Type Culture Collection (1 mentions) Acrylamide, by Bio-Rad (1 mentions) Penicillin streptomycin, by Euroclone (1 mentions) Temed, by Bio-Rad (1 mentions) Horse radish peroxidase linked anti rabbit secondary antibodies, by Cell Signaling Technology (1 mentions) Optiphase hisafe 2, by Beckman Coulter (1 mentions) Adenosine diphosphate (adp), by Merck Group (1 mentions) Dithiothreitol (dtt), by Merck Group (1 mentions) F 12k medium, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Lonza (1 mentions)

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ATCC CCL-185 CCL-185 CCL-185TM

11 protocols using atcc ccl 185tm

1

In Vitro Cultivation of Prostate and Lung Cancer Cell Lines

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Human prostate cancer (DU145: human epithelial cells- ATCC® HTB-81TM) purchased from ATCC (Manassas, VA, USA) and lung cancer (A549: human epithelial cells- ATCC® CCL-185TM) cells, kindly donated by Laboratory Medicine Department, RMIT University, were used in this study.
DU145 cells were cultured and maintained in MEM Alpha + GlutaMAXTM and 15 mM HEPES (Gibco®, Grand Island, NY, USA), 10% foetal bovine serum (FBS) (Gibco®) and 1% Penicillin-Streptomycin (Gibco®). A549 cells were cultured and maintained in DMEM/F12 supplemented with L-Glutamine and 15 mM HEPES (Gibco®), 10% FBS (Gibco®) and 1% Penicillin-Streptomycin (Gibco®).
Both cell lines initially were cultured and grown to 80% confluence in a 25 cm2 flask and then were sub-cultured. Incubation conditions during the experiments was 37 °C with 5% CO2 in a humidified environment.
Shahhoseini E., Feltis B.N., Nakayama M., Piva T.J., Pouniotis D., Alghamdi S.S, & Geso M. (2019). Combined Effects of Gold Nanoparticles and Ionizing Radiation on Human Prostate and Lung Cancer Cell Migration. International Journal of Molecular Sciences, 20(18), 4488.
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2

Lung Cancer Cell Culture and Irradiation

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A549 and H1299 non-small cell lung cancer cells21 (link),22 (link) and normal lung epithelial cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% FBS, 2 mM of L-glutamine, 100 U/mL of penicillin, and 100 μg/mL of streptomycin (Invitrogen) at 37 °C with 5% CO2 in a humidified incubator. Cell number was determined in duplicates using a haemocytometer. Irradiation was performed using a cobalt-60 gamma-ray irradiator (Theratron).
The A549 and H1299 cell lines were purchased from ATCC (ATCC® CCL-185TM and ATCC® CRL-5803TM, respectively), and all experiments were performed within 12 months. Details on the cell lines can be found at https://www.atcc.org/Products/All/CCL-185.aspx?slp=1#generalinformation and http://www.atcc.org/Products/All/CRL-5803.aspx#characteristics. The cell lines were authenticated by ATCC, and details and methods of characterization are available at http://www.atcc.org/Products/Cells_and_Microorganisms/Testing_and_Characterization/STR_Profiling_Analysis.aspx.
Both cell lines constitutively activated the NFκB pathway, as shown by the higher levels of pp65 expression in the nuclei of cells under standard conditions compared with normal human lung protein extracts (Figure S1).
Tsolou A., Liousia M., Kalamida D., Pouliliou S., Giatromanolaki A, & Koukourakis M. (2017). Inhibition of IKK-NFκB pathway sensitizes lung cancer cell lines to radiation. Cancer Biology & Medicine, 14(3), 293-301.
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3

Quantifying Influenza A Infection Efficiency

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A549 cells were obtained from ATCC (ATCC® CCL-185TM). A549 and 293FT cells were passaged and cultured in DMEM + l-glutamine, 10% FBS, 1% PenStrep at 37 degrees Celsius, and 5% CO2 in humidified conditions. PR8 and PR8ΔNS1 stocks were produced and titered as previously described [34 (link)]. Single-round infection assays on A549 cells was performed by infecting cells with PR8 or PR8ΔNS1. Cells were then fixed using the Foxp3/Transcription Factor Staining Buffer Set (Affymetrix, Santa Clara, CA, USA) according to the manufacturer’s protocol. Cells were stained for nucleoprotein using anti-influenza A virus nucleoprotein antibody [431] (FITC) (ab20921) (Abcam), then taken to the flow cytometer for measurement.
Razooky B.S., Obermayer B., O’May J.B, & Tarakhovsky A. (2017). Viral Infection Identifies Micropeptides Differentially Regulated in smORF-Containing lncRNAs. Genes, 8(8), 206.
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4

Characterization of Human Cancer Cell Lines

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American Type Culture Collection (ATCC) cell lines Human lung adenocarcinoma (A549; ATCC® CCL-185TM), breast adenocarcinoma (MDA-MB 231; ATCC® CRM-HTB-26TM) and lung embryonic fibroblast (MRC-5; ATCC® CCL-171TM) cell lines were obtained from ATCC. Human ovarian clear cell adenocarcinoma (RMG-1; CVCL-1662) cell line was obtained from ATCC and Japanese collection of research and bioresources (JCRB). All of the above mentioned cell lines were cultured in DMEM (Dulbecco's modified essential medium) growth media (Sigma, D6429) supplemented with 10% Fetal Bovine Serum (Sigma, F0804) and 1% penicillin/streptomycin (Sigma, P4333). MDA-MB 231 expresses mutant p53 while the others express wild type p53. All cultures were maintained at 37°C with 5% CO2.
Monchusi B, & Ntwasa M. (2017). Methyl pyruvate protects a normal lung fibroblast cell line from irinotecan-induced cell death: Potential use as adjunctive to chemotherapy. PLoS ONE, 12(8), e0182789.
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5

Enzymatic Activity Assay for NT5C2

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[8-14C]-IMP and [8-14C]-inosine were purchased from Moravek Biochemicals and Radiochemicals (Brea, CA, USA); DTT, ATP, ADP, AMP, MgCl2 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB), 3,3′,5,5′-tetramethylbenzidine (TMB), STOP solution, primary antibody against NT5C2 and 2-deoxyglucose (2-dG) were from Sigma (Milano, Italy) and polyethyleneimine (PEI)-cellulose precoated thin-layer plastic sheets were obtained from Merck (Darmstadt, Germany) and prewashed once with 10% NaCl and three times with deionized water before use. DE-81 chromatographic paper was from Whatman (Madstone, UK), horse radish peroxidase (HRP)-linked anti-rabbit secondary antibodies were from Cell Signaling (Danvers, MA, USA) and scintillation liquid Optiphase Hisafe 2 was from Beckman (Brea, CA, USA). Acrylamide and TEMED were from Bio-Rad (Segrate, Milan); penicillin/streptomycin; glutamine and fetal bovine serum (FBS), RPMI and DMEM media were from Euroclone (Pero, Milan) and a Duolink kit for the proximity ligation assay (PLA) assay was from Olink Bioscience (St. Louis, MO, USA). The human lung carcinoma (A549) and the human astrocytoma (ADF) cell lines were purchased from ATCC (ATCC® CCL-185TM) and routinely tested for Mycoplasma contamination by PCR. All other reagents were of reagent grade.
Pesi R., Allegrini S., Balestri F., Garcia-Gil M., Cividini F., Colombaioni L., Jordheim L.P., Camici M, & Tozzi M.G. (2021). Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic Regulator. Cells, 10(1), 182.
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6

Culturing KRAS-Mutant A549 Cancer Cells

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The human A549 KRAS-mutated cancer cell line was bought from the ATCC® Manassas, VI USA (ATCC CCL-185TM), and cultured in F­12 K medium (Life-Technologies® Carlsbad, CA USA), supplemented with 10% fetal bovine serum (Perbio®, Waltham, MA USA) and 1% penicillin/streptomycin (Invitrogen®, Carlsbad, CA USA) at 37 °C in a 5% CO2 atmosphere. Cultured cells were tested for mycoplasma regularly.
Massy E., Rousseau J., Gueye M., Bonnelye E., Brevet M., Chambard L., Duruisseaux M., Borel O., Roger C., Guelminger R., Pialat J.B., Gineyts E., Bouazza L., Millet M., Maury J., Clézardin P., Girard N, & Confavreux C.B. (2021). Serum total periostin is an independent marker of overall survival in bone metastases of lung adenocarcinoma. Journal of Bone Oncology, 29, 100364.
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7

Cytotoxicity Assay for P. aeruginosa

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The human A549 lung epithelial cell type II line (ATCC-CCL185TM) (American Type Culture Collection, Manassas, VA) was grown at 37°C in 5% CO2 atmosphere in Dulbecco’s modified Eagle’s medium (DMEM; Lonza) supplemented with 10% fetal calf serum (Lonza) and 1% of penicillin and streptomycin (Penistrep; Lonza). For cytotoxicity assays, cells were seeded in 24-well plates at a final density of 3 × 105 cells per well and grown for 48 h before use. A minimum of 24 h before infection assays, cells were deprived of antibiotics and fetal calf serum by the addition of a fresh serum- and antibiotic-free medium. For the assays, A549 cells were incubated for 2 h with control or pretreated P. aeruginosa PA14 at a multiplicity of infection of 10. Bacterial cytotoxicity was determined by measurement of lactate dehydrogenase (LDH) release. LDH is a stable cytosolic enzyme that diffuses into the culture medium upon cell lysis. This marker of cytotoxicity was assayed using the Cytotox 96 enzymatic assay (Promega, Charbonnières, France) as previously described (17 (link)).
Rosay T., Bazire A., Diaz S., Clamens T., Blier A.S., Mijouin L., Hoffmann B., Sergent J.A., Bouffartigues E., Boireau W., Vieillard J., Hulen C., Dufour A., Harmer N.J., Feuilloley M.G, & Lesouhaitier O. (2015). Pseudomonas aeruginosa Expresses a Functional Human Natriuretic Peptide Receptor Ortholog: Involvement in Biofilm Formation. mBio, 6(4), e01033-15.
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8

Culturing A549 Lung Cancer Cells

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A549 lung adenocarcinoma cell line (ATCC® CCL-185TM) was purchased from the American Type Culture Collection (ATCC® PCS-301-010™). The cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Sigma-Aldrich, (Taufkirchen, Germany) and supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, Taufkirchen, Germany) and 1% penicillin/streptomycin mixture (Pen/Strep, 10,000 IU/mL; Sigma-Aldrich, Taufkirchen, Germany) and further maintained in standard conditions (humidified atmosphere with 5% CO2 and 37 °C).
Kis B., Pavel I.Z., Haidu D., Ștefănuț M.N., Diaconeasa Z., Moacă E.A., Dehelean C.A., Șipos S., Ivan A, & Danciu C. (2021). Inorganic Element Determination of Romanian Populus nigra L. Buds Extract and In Vitro Antiproliferative and Pro-Apoptotic Evaluation on A549 Human Lung Cancer Cell Line. Pharmaceutics, 13(7), 986.
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9

Cell Culture of A549 Lung Cancer Cells

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The human lung cancer cell line A549 (ATCC® CCL-185TM) from American Type Culture Collection (ATCC) was used in this study. The Rosewell Park Memorial Institute 1640 medium (RPMI) with 1% antibiotics (amphotericin B and penicillin/streptomycin) and 10% fetal bovine serum (FBS) was used for cell culture. Cells were incubated at 37 °C in an 85% humified incubator with 5% carbon dioxide (CO2) to 90% confluence. Afterward, the used complete medium was discarded, followed by a thrice wash with Hank’s balanced salt solution (HBSS) (10 mL). Experimental cells were detached from the culture flasks with TrypLE (Invitrogen, 12605-028) (3 mL). After that, experimental cells were cultured in 3.5 cm diameter culture dishes containing 2 mL of the complete medium at a concentration of 5 × 105 cells/mL.
Chota A., George B.P, & Abrahamse H. (2022). In Vitro Cell Death Mechanisms Induced by Dicoma anomala Root Extract in Combination with ZnPcS4 Mediated-Photodynamic Therapy in A549 Lung Cancer Cells. Cells, 11(20), 3288.
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10

Sterile Cell Culture Techniques

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Human lung adenocarcinoma cells (A549, ATCC® CCL‐185TM) and human normal lung cells (CCD‐19Lu, ATCC® CCL‐210TM) from the American Type Culture Collection (ATCC) were used. Sterilisation of all liquids and plastic materials used in the experiments in an autoclave at 121°C, 1.5 atm for 20 min (Eryiğit A.Ş. Turkey), sterilisation of glass and metal materials at 180°C for 2 h in a dry air sterilizer (oven, Heraeus). Sterilisation of other liquid solutions was achieved by passing them through sterile disposable injector filters (Sartorius Stedim Biotech) with a pore diameter of 0.22 μm. Studies were conducted in aseptic conditions in a laminar flow cabinet (Mars Scanlaf, Labogene ApS, Denmark), protected from light.
Kurban B., Tuncel T., Görgülü Ş., Kar F., Öztürk A, & Özek T. (2023). Elemi essential oil nanocapsulated drug ameliorates lung cancer via oxidative stress, apoptosis and inflammation pathway. Journal of Cellular and Molecular Medicine, 27(13), 1887-1899.
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