Mcf 7

Manufactured by American Type Culture Collection

Sourced in United States, United Kingdom, China, Germany, France, Japan, Italy, Switzerland, Sweden, India, Canada, Egypt, Australia, Holy See (Vatican City State), Brazil

MCF-7 is a cell line derived from human breast adenocarcinoma. It is an adherent epithelial cell line that can be used for in vitro studies.

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5 027 protocols using mcf 7

Characterization of Breast Cancer Cell Lines

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All human breast cancer cell lines (MCF7, SKBR3, MDA-MB-231 and T47D) were purchased from American Type Culture Collection (ATCC, Rockville, MD). Stable COX-2 over expressing MCF7-COX-2 and SKBR3-COX-2 cell lines was generated in our laboratory as described previously^{21 (link)}. MCF7 and MCF7-COX-2 cells were grown in Eagle’s Minimum Essential Medium (EMEM), supplemented with 0.4 µl/mL human insulin (ATCC). SKBR3 and SKBR3-COX-2 were grown in McCoy’s 5 A Modified Medium with L-glutamine (GIBCO, ON). MDA-MB-231 (high COX-2, ER and HER-2 negative) and T47D (low COX-2, ER positive and HER-2 negative) were grown in RPMI-1640 minimal essential medium (GIBCO, ON). All media were supplemented with 10% fetal bovine serum (FBS) and 100 U/mL penicillin and 100 µg/mL streptomycin (Sigma, ON) and maintained in a humidified incubator with 5% CO2 at 37 °C. Additionally MCF7-COX-2 and SKBR3-COX-2 and their respective mock cell lines maintained with Geneticin® (GIBCO, ON) at 500 μg/mL.

Majumder M., Dunn L., Liu L., Hasan A., Vincent K., Brackstone M., Hess D, & Lala P.K. (2018). COX-2 induces oncogenic micro RNA miR655 in human breast cancer. Scientific Reports, 8, 327.

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Tamoxifen-Resistant MCF-7 Cell Line Development

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MCF-7 (ATCC HTB-22) cells were obtained from Beijing Zhongyuan Limited (official ATCC distributors; Beijing, China) and MCF-7-TAM were generated by exposing MCF-7 cells to increasing concentrations of tamoxifen over a period of 6 months starting with a dose equivalent to IC-5 of tamoxifen. Cells were maintained in the culture medium and passaged, as per ATCC’s instructions. All cells were cultured in 5% CO₂ humidified atmosphere at 37 °C within our laboratory. tamoxifen and 5-azacytidine were purchased from Sigma Chemical Company (Shanghai, China).

Wang Q., Gun M, & Hong X.Y. (2019). Induced Tamoxifen Resistance is Mediated by Increased Methylation of E-Cadherin in Estrogen Receptor-Expressing Breast Cancer Cells. Scientific Reports, 9, 14140.

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Generating Drug-Resistant Breast Cancer Cells

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Human BC cell lines MDA-MB-231, MCF-7, BT474, and HCC1937 (parental drug-sensitive cells) provided by American Type Culture Collection (ATCC, Manassas, VA, USA) were exposed to gradient concentrations of different drugs (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany), respectively, to generate cisplatin-resistant BT474 (BT474/Cis) cells, doxorubicin-resistant MCF-7 (MCF-7/Dox) cells, and fulvestrant-resistant HCC1937 (HCC1937/Ful) cells. Cells were cultivated in complete modified Eagle's medium with 10% fetal bovine serum.

Wang B., Wang Y., Wang X., Gu J., Wu W., Wu H., Wang Q, & Zhou D. (2022). Extracellular Vesicles Carrying miR-887-3p Promote Breast Cancer Cell Drug Resistance by Targeting BTBD7 and Activating the Notch1/Hes1 Signaling Pathway. Disease Markers, 2022, 5762686.

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Breast Cancer Cell Maintenance Protocol

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MCF-7 and MDA-MB-231 were purchased from
the American Type Culture Collection (Rockville, MD, USA) and maintained
in E-MEM (MCF-7) or DMEM (MDA-MB-231) supplemented with 10% fetal
bovine serum (FBS), 2 mM l-glutamine, 500 U/mL penicillin,
and 50 μg/mL streptomycin. The MCF-7TAM cell line was established
by growing MCF-7 cells in MEM medium (without phenol red and with
charcoal-treated 10% FBS) containing 10^–7 M 4-OH-tamoxifen
as already described.^{23 (link),24 (link),44 (link)} MCF10A was purchased from the American Type Culture Collection (Rockville,
MD, USA) and maintained in low glucose DMEM medium, supplemented with
20% fetal bovine serum (FBS), 2 mM l-glutamine, 500 U/mL
penicillin, 50 μg/mL streptomycin, insulin (25 U), hydrocortisone
(0.5 μg/mL), and EGF (5 nM).

Farabegoli F., Granja A., Magalhães J., Purgato S., Voltattorni M, & Pinheiro M. (2022). Epigallocatechin-3-gallate Delivered in Nanoparticles Increases Cytotoxicity in Three Breast Carcinoma Cell Lines. ACS Omega, 7(46), 41872-41881.

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Culturing Diverse Cell Lines with FBS

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Cells (MCF‐10A, MCF‐7, MDA‐MB‐231, and 293T) were purchased from the American Type Culture Collection (ATCC, Manassas, Virginia, USA). MCF‐10A cells were cultured in mammary epithelial growth medium (MEGM) (Catalog No. CC‐3150). MCF‐7 were cultured in Eagle's Minimum Essential Medium (EMEM) (ATCC, 30‐2003), with MDA‐MB‐231 in Roswell Park Memorial Institute‐1640 solution (RPMI) (Solarbio, Catalog No. 3180) and 293T in Dulbecco's modified Eagle’s medium (DMEM) (Solarbio, Catalog No.11995).
All four kinds of cells were cultured at 37°C with 5% CO₂, and their media was supplemented with 10% fetal bovine serum (FBS).

Zhang L., Liu Q., Mu Q., Zhou D., Li H., Zhang B, & Yin C. (2020). MiR‐429 suppresses proliferation and invasion of breast cancer via inhibiting the Wnt/β‐catenin signaling pathway. Thoracic Cancer, 11(11), 3126-3138.

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Culturing Diverse Cancer Cell Lines

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Breast cancer (MDA-MB231, MCF-7) and colon cancer (LS180, SW480) cell lines were obtained from the ATCC (Manassas, VA). The pancreatic cancer cell lines (PATU8988T, Panc1) were kindly provided by Dr. A. Kimmelman (Harvard Medical School, Boston, MA). The T2 cell line was provided by Dr. J. Molldrem (University of Texas M. D. Anderson Cancer Center, Houston, TX). MDA-MB231 and SW480 cell lines were cultured in Leibovitz’s L-15 Medium (ATCC); MCF-7 and LS180 cell lines were cultured in EMEM (Gibco-Life Technologies, Rockville, MD); and Panc1, PATU8902, and T2 cell lines were cultured in DMEM (Gibco-Life Technologies) media. All media were supplemented with 10% fetal calf serum (FCS; BioWhittaker, Walkersville, MD), 100 IU/ml penicillin, and 100 μg/ml streptomycin (Gibco-Life Technologies).

Bae J., Keskin D.B., Cowens K., Lee A.H., Dranoff G., Munshi N.C, & Anderson K.C. (2015). Lenalidomide Polarizes Th1-specific Anti-tumor Immune Response and Expands XBP1 Antigen-Specific Central Memory CD3+CD8+ T cells against Various Solid Tumors. Journal of leukemia (Los Angeles, Calif.), 3(2), 178.

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Breast Cancer Cell Line Characterization

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MCF-7 (ER+/PR+/HER2−, BRCA1 proficient), MDA-MB-231 (ER−/PR−/HER2−, BRCA1 proficient), MDA-MB-468 (ER−/PR−/HER2−, BRCA1 proficient) and MDA-MB-436 (ER−/PR−/HER2−, BRCA1 deficient) were purchased from ATCC and were grown in RPMI (MCF-7) or DMEM (MDA-MB-231, MDA-MB-468 and MDA-MB-436) medium with the addition of 10% foetal bovine serum and 1% penicillin/streptomycin. Cells in culture were routinely checked for mycoplasma contamination by PCR (Sigma, catalog no. MP0035).The cells characterisation were performed by ATCC and passaged in the laboratory for fewer than 6 months.

Arora A., Parvathaneni S., Aleskandarany M.A., Agarwal D., Ali R., Abdel-Fatah T., Green A.R., Ball G.R., Rakha E.A., Ellis I.O., Sharma S, & Madhusudan S. (2016). Clinicopathological and functional significance of RECQL1 helicase in sporadic breast cancers. Molecular cancer therapeutics, 16(1), 239-250.

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Murine and Human Breast Cancer Cell Lines

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Murine 4T1, a 6-Thioguanine resistant cell line, human MCF-7 and human MDA-MB-231 cell lines were obtained from American Type Cell Collection (ATCC; Chicago, IL, USA). ATCC authenticated the human cell lines by using short tandem repeat profiling and the mice cell line was confirmed to be from mice by cytochrome C oxidase 1 gene assay. MCF-7 cells were cultured in MEM, 4T1 and MDA-MB-231 in RPMI-1640, media containing FBS (10 %, v/v) (ATCC), penicillin (100 µU/ml), streptomycin (100 µg/ml) (Sigma-Aldrich, Oakville, ON) at 37 °C in a humidified atmosphere with 5 % CO₂.

Vuong T., Mallet J.F., Ouzounova M., Rahbar S., Hernandez-Vargas H., Herceg Z, & Matar C. (2016). Role of a polyphenol-enriched preparation on chemoprevention of mammary carcinoma through cancer stem cells and inflammatory pathways modulation. Journal of Translational Medicine, 14, 13.

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Cell Line Characterization and Cultivation

Cited 1 time

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MDA-MB-231 (HTB-26), MDA-MB-468 (HTB-132), MCF10A (CRL-10317), and MCF-7 (HTB-22) were purchased from ATCC in 2010 (MCF-7), 2012 (MDA-MB-468), and 2014 (MDA-MB-231 and MCF10A). Primary human fibroblast cells were kindly provided by Kathrin Scheckenbach, Düsseldorf, Germany in 2014. TMD231 cells were obtained from Dr. Harikrishna Nakshatri in 2009 and are a derivative of the MDA-MB-231 cell line (19 (link)). TMD231 cells were transduced with E2-Crimson lentiviral vector, p2CL7CR2wo, (TMD231-CR) for in vivo imaging as described (20 (link)). Upon receipt, cell stocks were cryopreserved at low passage. Authentication of molecular profiles was verified by short tandem repeat (STR) analysis (IDEXX BioResearch), and cells tested negative for mycoplasma. TMD231 cells were cultured in MEM-α (Gibco) supplemented with 10% FBS (Atlanta Biologicals) and 1% HEPES (Invitrogen). MDA-MB-231, MDA-MB-468, and primary human fibroblast cells were cultured in DMEM (Gibco) supplemented with 10% FBS (Atlanta Biologicals). MCF10A cells were cultured in Medium 171 (Gibco) supplemented with 1% MEGS (Invitrogen) and 0.1% cholera toxin (Sigma). All cells were cultured at 37°C with 5% CO₂.

Tonsing-Carter E., Bailey B.J., Saadatzadeh M.R., Ding J., Wang H., Sinn A.L., Peterman K.M., Spragins T.K., Silver J.M., Sprouse A.A., Georgiadis T.M., Gunter T.Z., Long E.C., Minto R.E., Marchal C.C., Batuello C.N., Safa A.R., Hanenberg H., Territo P.R., Sandusky G.E., Mayo L.D., Eischen C.M., Shannon H.E, & Pollok K.E. (2015). Potentiation of carboplatin-mediated DNA damage by the Mdm2 modulator Nutlin-3a in a humanized orthotopic breast-to-lung metastatic model. Molecular cancer therapeutics, 14(12), 2850-2863.

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Culturing Breast Cell Lines for Research

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Breast cancer cell lines MCF-7, MDA-MB-231 and HCC1937, as well as normal breast cell line MCF 10A were all obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). MCF-7 cells were cultured in Eagle’s Minimum Essential Medium (No. 30-2003; ATCC, Manassas, VA, USA), supplemented with 0.01 mg/mL human recombinant insulin and 10% fetal bovine serum (FBS; Gibco, MA, USA). HCC1937 cells were grown in ATCC-formulated RPMI-1640 Medium (No. 30-2001) filling with 10% FBS. MDA-MB-231 cells were cultured in ATCC-formulated Leibovitz’s L-15 Medium (No. 30-2008) filling with 10% FBS. MCF 10A were cultured in MEGM Kit (No. CC-3150; Lonza/Clonetics Corporation, CA, USA) with 100 ng/mL cholera toxin (No. C8052; Sigma, MA, USA). All cells were kept in a humidified atmosphere at 37 °C with 5% CO₂.

Zhang G., Zhong L., Luo H, & Wang S. (2019). MicroRNA-155-3p promotes breast cancer progression through down-regulating CADM1. OncoTargets and therapy, 12, 7993-8002.

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