A2780cis cell

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A2780cis cells are a type of ovarian cancer cell line commonly used in research. They are a subline of the parental A2780 cell line and are known to be resistant to the chemotherapeutic drug cisplatin. A2780cis cells are frequently utilized in studies related to drug resistance and cancer biology.

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A2780cis (39 citations) A2780CIS cell line (2 citations)

7 protocols using a2780cis cell

Cell Line Cultivation and Characterization

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HeLa, Saos-2, HS-5, HepG2, T98G, and A2780 cells were purchased from American-type Culture Collection (ATCC, USA), A2780cis cell from Sigma, and Kuramochi and OVSAHO cell lines from the lab of Dinulescu lab at Harvard Medical School. Dulbecco’s modified Eagle’s medium (DMEM), McCoy’s 5a medium, and 1640 Medium were purchased from ATCC, and fetal bovine serum (FBS) and penicillin/streptomycin from Gibco by Life Technologies. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from ACROS Organics, ER stress antibody kit from Cell Signaling Technology, and other antibodies from Abcam.

Wang H., Feng Z., Yang C., Liu J., Medina J.E., Aghvami S.A., Dinulescu D.M., Liu J., Fraden S, & Xu B. (2018). Unraveling the Cellular Mechanism of Assembling Cholesterols for Selective Cancer Cell Death. Molecular cancer research : MCR, 17(4), 907-917.

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Cell Line Authentication and NAC Treatment Protocol

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PEO1, PEO4, and A2780CIS cell lines were purchased from SIGMA (St. Louis, MO, USA). Lenti-X 293T cell line was purchased from Takara Bio (Kusatsu, Shiga, Japan). A2780CIS cells were cultured in RPMI-1640 medium supplemented with 10% FBS and 100 units/mL of penicillin and 100 µg/mL streptomycin. PEO1 and PEO4 cells were cultured in RPMI-1640 medium supplemented with 2 mM glutamine, 2 mM sodium pyruvate, and the same concentrations of FBS and penicillin/streptomycin as described above. All cells were maintained in an incubator at 37 °C with 5% CO₂. All cell lines were authenticated before experimentation and tested negative for mycoplasma.
For experiments with N-Acetyl-L-cysteine (NAC; Sigma, St. Louis, MO, USA), cells were pre-treated with 2.5 mM NAC thirty minutes prior to saline or cisplatin treatment.

Heiserman J.P., Minhas Z., Nikpayam E, & Cheon D.J. (2023). Targeting Heat Shock Protein 27 and Fatty Acid Oxidation Augments Cisplatin Treatment in Cisplatin-Resistant Ovarian Cancer Cell Lines. International Journal of Molecular Sciences, 24(16), 12638.

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Ovarian Cancer Cell Line Culturing

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A2780 and A2780cis cells were purchased from Sigma-Aldrich (St. Louis, Missouri, US), SKOV-3 and OVCAR-3 cells were purchased from the American Type Culture Collection (Manassas, Virginia, US) and SKOV-3cis and OVCAR-3cis were provided by Axis Bioservices, (Coleraine, UK). A2780 and A2780cis cells were cultured with RPMI 1640 supplemented with 10% Fetal Bovine Serum (FBS), and 1% penicillin-streptomycin (all from Gibco by ThermoFisher, Waltham, Massachusetts, US). OVCAR-3 and OVCAR-3cis cells were cultured with RPMI 1640 supplemented with 20% FBS, 1% penicillin-streptomycin and human insulin (Sigma-Aldrich) to a final concentration of 10 µg/ml. SKOV-3 and SKOV-3cis cells were cultured with McCoys 5A Medium + L-glutamine (Gibco by ThermoFisher) supplemented with 10% FBS, and 1% penicillin-streptomycin. During alternate cell passages, media for cisplatin-resistant cell lines was additionally supplemented with cisplatin at final concentrations of 3 µM for SKOV-3cis, 1.5 µM for OVCAR-3cis and 1 µM for A2780cis cells, respectively.

Howard D., James D., Garcia-Parra J., Pan-Castillo B., Worthington J., Williams N., Coombes Z., Rees S.C., Lutchman-Singh K., Francis L.W., Rees P., Margarit L., Conlan R.S, & Gonzalez D. (2022). Dinaciclib as an effective pan-cyclin dependent kinase inhibitor in platinum resistant ovarian cancer. Frontiers in Oncology, 12, 1014280.

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Transfection of A2780cis Cells with siRNA

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The day before transfection, A2780cis cells (Sigma-Aldrich) were seeded at ∼5–7 × 10⁴ cells per well of a 96-well plate (Perkin Elmer) in 100 μl of RPMI with 10% FBS without antibiotics. For each transfection well, Silencer Select Pre-designed siRNA (Ambion Life Technologies, Grand Island, NY) was diluted into 25 μl Opti-MEM reduced serum medium for final concentration of 10 nm per well. In a separate tube, 0.5 μl of lipofectamine RNAiMAX transfection reagent was added per well and diluted into 25 μl Opti-MEM reduced serum medium and mix. After a 5-min incubation the tubes were combined then incubated at room temperature for 30 min to allow siRNA-liposome complexes to form. Once complexed, the mix was added to cells (growth media removed), and then diluted with growth media to 100 μl final volume. Cells are then incubated with siRNA for 18 h at 37 °C in a CO₂ incubator, at which time the mixture was removed and replaced with complete growth medium. Cells were screened for Fab binding after 48 h of siRNA addition.

Hornsby M., Paduch M., Miersch S., Sääf A., Matsuguchi T., Lee B., Wypisniak K., Doak A., King D., Usatyuk S., Perry K., Lu V., Thomas W., Luke J., Goodman J., Hoey R.J., Lai D., Griffin C., Li Z., Vizeacoumar F.J., Dong D., Campbell E., Anderson S., Zhong N., Gräslund S., Koide S., Moffat J., Sidhu S., Kossiakoff A, & Wells J. (2015). A High Through-put Platform for Recombinant Antibodies to Folded Proteins. Molecular & Cellular Proteomics : MCP, 14(10), 2833-2847.

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Cultured Human Ovarian Cancer Cell Lines

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Human ovarian cancer cisplatin-sensitive cell line A2780 cells (Sigma-Aldrich, cat. no. 93112519) and cisplatin-resistant A2780-cis cells (Sigma-Aldrich, cat. no. 93112517) were cultured in T-175 tissue culture flasks with 30 ml growth medium in a humidified atmosphere of 5% CO₂ at 37°C. Growth medium was made with RPMI 1640 Medium (GIBCO, USA) with 10% fetal bovine serum (FBS). Growth medium was replaced every other day and cells were passaged at 75% confluence.

Sima N., Sun W., Gorshkov K., Shen M., Huang W., Zhu W., Xie X., Zheng W, & Cheng X. (2018). Small Molecules Identified from a Quantitative Drug Combinational Screen Resensitize Cisplatin's Response in Drug-Resistant Ovarian Cancer Cells. Translational Oncology, 11(4), 1053-1064.

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Cell Line Authentication and Culture

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HeLa, T98G, HepG-2, HS-5 and Saos-2 cell lines were purchased from ATCC between 2010 and 2017. A2780cis cells were obtained from Sigma-aldrich in 2016. Kuramochi and OVSAHO were kindly provided by Prof. Dinulescu (Harvard medical school). All cell lines were authenticated using short tandem repeat DNA fingerprinting. A2780cis cells were cultured in RPMI 1640 medium supplemented with 10% v/v fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin (cisplatin only necessary every 2–3 passages). HeLa cells, T98G, and HepG-2 cells were cultured in MEM medium supplemented with 10% v/v fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin; HS-5 cells were cultured in Dulbecco’s Modified Eagle’s medium (DMEM) supplemented with 10% v fetal bovine serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin; Saos-2 cells were cultured in McCoy’s 5a medium (for Saos-2) supplemented with 15% v/v fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin; Kuramochi and OVSAHO cell lines were cultured in RPMI-1640 medium with 10% FBS and 1% P/S. All cells were incubated at 37 °C in a humidified atmosphere of 5% CO₂.

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Cell Line Culture and Authentication Protocol

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A2780 and A2780CIS cells were purchased from SIGMA (St. Louis, MO, USA). ES2, SKOV3, and A204 cells were purchased from ATCC (Manassas, VA, USA). Lenti-X 293T cells were purchased from Takara bio (formerly Clontech, Kusatsu, Shiga, Japan). A2780 and A2780CIS cells were cultured in RPMI (Gibco Life Technologies; Waltham, MA, USA) supplemented with 10% FBS and 1× penicillin/streptomycin. ES2, SKOV3, A204, and Lenti-X 293T cell lines were cultured in DMEM (Gibco Life Technologies; Waltham, MA, USA) supplemented with 10% FBS (Sigma-Aldrich, St. Louis, MO, USA) and 1× penicillin/streptomycin (Gibco Life Technologies; Waltham, MA, USA). All cells were cultured at 37 °C with 5% CO₂. All cell lines tested negative for mycoplasma contamination and were authenticated before experimentation.

Heiserman J.P., Nallanthighal S., Gifford C.C., Graham K., Samarakoon R., Gao C., Sage J.J., Zhang W., Higgins P.J, & Cheon D.J. (2021). Heat Shock Protein 27, a Novel Downstream Target of Collagen Type XI alpha 1, Synergizes with Fatty Acid Oxidation to Confer Cisplatin Resistance in Ovarian Cancer Cells. Cancers, 13(19), 4855.

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