Hcc1954 cells

Manufactured by American Type Culture Collection

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HCC1954 cells are a human breast cancer cell line derived from a primary ductal breast carcinoma. The cells are adherent and grow as a monolayer in culture. HCC1954 cells express the HER2/neu receptor and are used as a model system for studying HER2-positive breast cancer.

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HCC1954 (183 citations) HCC1954 cell line (3 citations)

8 protocols using hcc1954 cells

Cell Culture Protocols for Cell Lines

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HEK293T (ATCC CRL-3216) and U2OS (ATCC-HTB-96) were maintained in Dulbecco's Modified Eagle's Medium plus GlutaMax (ThermoFisher) supplemented with 10% (v/v) fetal bovine serum (FBS), at 37 °C with 5% CO₂. HCC1954 cells (ATCC CRL-2338) were maintained in RPMI-1640 medium (ThermoFisher Scientific) supplemented as described above. Immortalized rat astrocytes containing the ApoE4 isoform of the APOE gene (Taconic Biosciences) were cultured in Dulbecco's Modified Eagle's Medium plus GlutaMax (ThermoFisher Scientific) supplemented with 10% (v/v) fetal bovine serum (FBS) and 200 μg/mL Geneticin (ThermoFisher Scientific).

Komor A.C., Kim Y.B., Packer M.S., Zuris J.A, & Liu D.R. (2016). Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature, 533(7603), 420-424.

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Culturing Human Microvascular Endothelial Cells

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Human Microvascular Endothelial Cells (HMEC-1) were obtained from the Center for Disease Control and Prevention, Atlanta USA (Ades et al., 1992 (link)) and maintained in MCDB 131 medium containing 10% FCS, 10 mM L-Glutamine, 100 U/ml penicillin and 100 µg/ml streptomycin (All from Life Technologies, USA), 50 nM hydrocortisone (Sigma-Aldrich, USA), and 10 ng/ml rhEGF (Invitrogen, USA) up to passage 27 at 37°C, 5% CO₂ and ambient oxygen level (20%). Immortalized human mammary epithelial cells (HMLE) were kindly provided by dr. R.A. Weinberg and were cultured in human mammary epithelial growth media (cc-3150, Lonza) as described previously (Vervoort et al., 2013a (link)). MDA-MB-231 cells and HCC1954 cells were purchased from ATCC and cultured as described previously (Bruna et al., 2012 (link)). All cells were regularly checked for mycoplasma during the course of this study and only mycoplasma-negative cells have been used for the experiments described in this study.

Vervoort S.J., de Jong O.G., Roukens M.G., Frederiks C.L., Vermeulen J.F., Lourenço A.R., Bella L., Vidakovic A.T., Sandoval J.L., Moelans C., van Amersfoort M., Dallman M.J., Bruna A., Caldas C., Nieuwenhuis E., van der Wall E., Derksen P., van Diest P., Verhaar M.C., Lam E.W., Mokry M, & Coffer P.J. (2018). Global transcriptional analysis identifies a novel role for SOX4 in tumor-induced angiogenesis. eLife, 7, e27706.

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Cell Culture Protocols for Cell Lines

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Komor A.C., Kim Y.B., Packer M.S., Zuris J.A, & Liu D.R. (2016). Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature, 533(7603), 420-424.

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Culturing HCC1954 Cells on Scaffolds

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HCC1954 cells (derived from human ductal carcinoma cells, primary tumor), purchased from ATCC (Manassas, VA, USA) (https://www.atcc.org, accessed on 1 September 2023), are cultured in RPMI medium supplemented with 1% penicillin/streptomycin, 1% L-glutamine, and 10% FBS at 37 °C in a humified atmosphere with 5% CO₂ (SteriCult CO₂ incubator, Thermo Electron Corporation, Thermo Fisher Scientific, Cincinnati, OH, USA). Cells (1.8 × 10⁵) are seeded on hydrated scaffolds previously put into each well of an 8-well cell chamber slide.

Sieni E., Dettin M., Zamuner A., Conconi M.T., Bazzolo B., Balducci C., Di Barba P., Forzan M., Lamberti P, & Mognaschi M.E. (2023). Finite Element Evaluation of the Electric Field Distribution in a Non-Homogeneous Environment. Bioengineering, 10(9), 1062.

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Establishment of HCC1954 Cell Culture

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HCC1954 cells were obtained from ATCC (Manassas, VA). The cells were cultured and maintained in the laboratory in RPMI medium (Corning; Tewksbury, MA) supplemented with 10% FBS (Hyclone, Marlborough, MA) and l-glutamine (Sigma; St. Louis, MO) according to ATCC specifications [24 (link)]. Tissue extraction from tumor specimens and primary cell culture was carried out as described previously [24 (link), 25 (link)]. Briefly tissue was minced to less than 2 mm length, enzymatically digested, and plated onto collagen-fibronectin coated 4-well culture plates where cells were typically cultured on average for less than fourteen days before transfer to the analytical test procedure.

MacNeil I.A., Khan S.A., Sen A., Soltani S.M., Burns D.J., Sullivan B.F, & Laing L.G. (2022). Functional signaling test identifies HER2 negative breast cancer patients who may benefit from c-Met and pan-HER combination therapy. Cell Communication and Signaling : CCS, 20, 4.

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Evaluating ADC Cytotoxicity on Cell Lines

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HCC1954 cells (ATCC) and RAW-dual cells (Invivogen) were cultured according to the manufacturer’s instructions in high-glucose DMEM (Corning) supplemented with 10% FBS. Cells were pre-mixed (1:1) giving a final seeding density of 0.2 million cells per milliliter each. The cell mixture (90 μL) was seeded into a 96-well plate and treated with 10 μL of the appropriate test article (ADC, mAb, or PBS). The final concentration of test ADCs was 30, 6, 1.2, 0.24, and 0 μg/mL. The naked mAb control was administered at 30 μg/mL. The cells were cultured for 24 h at which point the supernatant was separated via centrifugation, and the activation of the NFκB pathway was determined using the QuantiBlue assay described above.

Fang S., Brems B.M., Olawode E.O., Miller J.T., Brooks T.A, & Tumey L.N. (2022). Design and Characterization of Immune-Stimulating Imidazo[4,5-c]quinoline Antibody-Drug Conjugates. Molecular pharmaceutics, 19(9), 3228-3241.

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ADC Cytokine Induction in Tumor-Immune Cells

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HCC1954 cells (ATCC) and hPBMCs (iXCells Biotechnologies) were pre-mixed in RPMI1640 media (Lonzo) supplemented with 10% FBS before seeding. The final seeding densities of HCC1954 cells and hPBMCs were 0.2 million per milliliter and 2 million cells per milliliter, respectively. The cell mixture was seeded to a 96-well plate with a seeding volume of 90 μL with 10 μL of the appropriate test article (ADC, mAb, or PBS). The final concentration of test ADCs was 30, 6, 1.2, 0.24, and 0 μg/mL. The naked mAb control was administered at 30 μg/mL. The cells were cultured for 24 h, and then the supernatants were separated via centrifugation. IFNα levels in supernatant samples were quantified using a human IFNα all subtype ELISA kit (Pbl assay science, Cat# 411351).

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Generating Genetically Modified HCC1954 Cells

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HCC1954 cells were purchased from ATCC (Manassas, Virginia) and maintained in 10% FBS supplemented RPMI1640 (R8758, Sigma, St. Louis, MO) and 2 μg/mL puromycin (Sigma) was added to HCC1954 pLKO.1 and shE2F3 cells. Lentiviral particles carrying shRNA against E2F3 were purchased from Open Biosystems (clone ID: TRCN0000013807). Both 2 μg/mL puromycin and 50 μg/mL hygromycin (H0654, Sigma) were added to grow HCC1954 shE2F3;GFP-Nek2 cells [18 ]. shE2F3 cells overexpressing GFP were generated as follows: the GFP expressing vector, pMONO-Hygro-GFP (Cat#pmonoh-GFP, InvivoGen, San Diego, CA) was transfected into shE2F3 cells and underwent hygromycin selection 48 hr after transfection. Pools of clones were harvested, GFP expression was confirmed and cells maintained in the same media as shE2F3;GFP-Nek2 cells.

Lee M., Oprea-Ilies G, & Saavedra H.I. (2015). Silencing of E2F3 suppresses tumor growth of Her2+ breast cancer cells by restricting mitosis. Oncotarget, 6(35), 37316-37334.

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