Mda mb 468

Manufactured by American Type Culture Collection

Sourced in United States, United Kingdom, China, Germany

MDA-MB-468 is a human breast cancer cell line derived from the pleural effusion of a 51-year-old female patient with metastatic breast adenocarcinoma. This cell line is commonly used in cancer research to investigate aspects of breast cancer biology.

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1 280 protocols using mda mb 468

Expansion and Maintenance of Breast Cancer Cell Lines

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The MCF-7, HCC1937, MDA-MB-231, MDA-MB-468, MDA-MB-453, and SK-BR-3 cell lines were acquired from American Type Culture Collection (Virginia, USA). The MDA-MB-468 with STAT3 overexpression cell line was generously provided by Dr. Liu CY, working in Division of Hematology and Oncology, Department of Medicine, Taipei Veterans General Hospital (Taipei, Taiwan). All cell lines were immediately expanded and frozen down immediately after acquiring. All cell lines could be restarted every 3 months from a frozen vial of the same batch of cells. Cells except for MDA-MB-468 with STAT3 overexpression were maintained as described culture medium by ATCC; MDA-MB-468 with STAT3 overexpression cells was maintained in L-15 medium with G418 700 μg/mL. All media were supplemented with 10% FBS (Caisson, USA), 100 units/mL penicillin, 100 mg/mL streptomycin, and 25 mg/mL amphotericin B (Caisson, USA). All human breast cancer cell lines were incubated in a humidified incubator at 37°C in an atmosphere of 5% CO₂ in air.

Chiu Y.H., Lee Y.Y., Huang K.C., Liu C.C, & Lin C.S. (2019). Dovitinib Triggers Apoptosis and Autophagic Cell Death by Targeting SHP-1/p-STAT3 Signaling in Human Breast Cancers. Journal of Oncology, 2019, 2024648.

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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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Cell Line Characterization and Cultivation

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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 Diverse Breast Cancer Cell Lines

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White (HCC1143 and MDA-MB-231) and African American (HCC1906 and MDA-MB-468) cell lines were purchased from ATCC. MDA-MB-231 and MDA-MB-468 cells were cultured in Leibovitz’s L-15 medium with 10% fetal bovine serum (ATCC) at 37 °C in 100% air. HCC1806 and HCC1143 cell lines were cultured in Roswell Park Memorial Institute medium with 10% fetal bovine serum (ATCC) at 37 °C and 5% CO₂.

Ogden A., Garlapati C., Li X.(., Turaga R.C., Oprea-Ilies G., Wright N., Bhattarai S., Mittal K., Wetherilt C.S., Krishnamurti U., Reid M.D., Jones M., Gupta M., Osan R., Pattni S., Riaz A., Klimov S., Rao A., Cantuaria G., Rida P.C, & Aneja R. (2017). Multi-institutional study of nuclear KIFC1 as a biomarker of poor prognosis in African American women with triple-negative breast cancer. Scientific Reports, 7, 42289.

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Culturing Human Breast Cancer Cell Lines

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Human breast cancer MDA-MB-231, BT-549, MDA-MB-468, BT-474, BT-20 and MCF-7 cell lines (American Type Culture Collection, ATCC, Manassas, VA) were grown in RPMI-1640 (Invitrogen, Carlsbad, CA, USA) (MDA-MB-231, BT-549 and MDA-MB-468), ATCC HybriCare Medium (BT-474) or Dulbecco’s modified Eagle’s medium (Invitrogen) (BT-20 and MCF-7) supplemented with 10% fetal bovine serum (FBS) (Invitrogen), in 95% air/5% CO2 atmosphere at 37 °C. Growth conditions for mouse NIH3T3 cells were previously reported26 (link).

Camorani S., Crescenzi E., Gramanzini M., Fedele M., Zannetti A, & Cerchia L. (2017). Aptamer-mediated impairment of EGFR-integrin αvβ3 complex inhibits vasculogenic mimicry and growth of triple-negative breast cancers. Scientific Reports, 7, 46659.

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Breast Cancer Cell Lines Cultivation

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HER-2^pos SKBR-3, triple negative MDA-MD-468, HER-2^pos, trastuzumab resistant HCC1419, and HER-2^pos, trastuzumab and lapatinib resistant MDA-MB-453 cell lines were obtained from American Type Culture Collection (ATCC; Rockwell, MA). MDA-MB-468, MDA-MB-453 and HCC1419 cell lines were re-authenticated by ATCC using STR prior to submission. Assays preformed using the SKBR-3 cell line were done within 6 months after the time of purchase from ATCC. Murine TUBO cells were a kind gift from Dr. Wei Zen Wei, (Wayne State University) and were derived from a neu transgenic mouse [45 (link)] and were cultured in RPMI medium. SKBR-3 cells were cultured in McCoy’s 5A Media (Corning, Manassas, VA), MDA-MB-468, MDA-MB-453, and HCC1419 cells were cultured in RPMI-1640 (ATCC). All media was supplemented with 10% v/v fetal calf serum (FBS; Atlanta Biologicals, Flowery Branch, GA), 100 units/ml of penicillin and 100 μg/ml of streptomycin sulfate (BioWhittaker, Walkersville, MD), 2 mmol/L glutamine (BioWhittaker), 1 mmol/L sodium pyruvate (BioWhittaker), and 1% non-essential amino acids (BioWhittaker). All cells were maintained in culture at 37°C in 5% CO₂ in a humidified atmosphere.

Showalter L., Czerniecki B.J, & Koski G.K. (2020). Th1 cytokines in conjunction with pharmacological Akt inhibition potentiate apoptosis of breast cancer cells in vitro and suppress tumor growth in vivo. Oncotarget, 11(30), 2873-2888.

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Cell Line Characterization and Cultivation

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The HMLE^HRASV12 cell line (H^RAS cells), obtained after transforming human mammary epithelial (HMLE) cells with HRAS T24 oncogene, was collected in 2011 from Robert Weinberg (Whitehead Institute, Cambridge, MA, USA), and cultured in DMEM-F12 medium (ATCC) containing 10% fetal bovine serum (FBS) from ATCC. The human breast cancer cell lines MDA-MB-231, HCC1806 and MDA-MB-468 were obtained from ATCC in 2015. ATCC uses the Promega PowerPlex 1.2 system and the Applied Biosystems Genotyper 2.0 software for amplicon analysis. We have not performed any further testing in our lab. MDA-MB-231 and MDA-MB-468 cells were propagated in Leibovitz L-15 medium (ATCC) containing 10% FBS. HCC 1806 cells were propagated in RPMI-1640 media (ATCC) supplemented with 10% FBS, 100 U/ml penicillin, and 100 μg/ml streptomycin. All cell lines were incubated at 37°C in a humidified atmosphere containing 5% CO₂, with the exception of the MDA-MB-231 and MDA-MB-468 cells that were grown in the Leibovitz L-15 medium formulation at 37°C in a free gas exchange with 100% atmospheric air.

Howard C.B., Mcdowell R., Feleke K., Deer E., Stamps S., Thames E., Singh V, & Pervin S. (2016). Chemotherapeutic Vulnerability of Triple-negative Breast Cancer Cell-derived Tumors to Pretreatment with Vernonia amygdalina Aqueous Extracts. Anticancer research, 36(8), 3933-3943.

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Cisplatin Effects on TNBC Cell Lines

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The normal human breast epithelial cell line MCF10A and the human TNBC cell lines HCC1937, BT-549, MDA-MB-231, MDA-MB-436, and MDA-MB-468 were purchased from ATCC (Manassas, VA, USA). MCF10A cells were cultured in DMEM/F12 medium supplemented with 5% heat-inactivated horse serum, 0.5 mg/mL hydrocortisone, 20 ng/mL EGF, 10 μg/mL insulin, 100 ng/mL cholera toxin, 100 IU/mL penicillin, and 100 μg/mL streptomycin. HCC1937 cells were maintained in RPMI 1640 medium (Life Technologies, Carlsbad, CA, USA). BT-549 cells were cultured in RPMI 1640 medium with 10% fetal bovine serum (FBS) and 10 μg/mL insulin. MDA-MB-231 and MDA-MB-468 cells were cultured in ATCC-formulated Leibovitz’s L-15 medium supplemented with 10% FBS, while MDA-MB-436 cells were grown in the same medium with 10 μg/mL insulin and 16 μg/mL glutathione. All the cells were maintained in a humidified incubator at 37°C with 5% CO₂. Cells were passaged by trypsinization after reaching approximately 80%–90% confluence. The detached cells were seeded into a new flask at a density of 2.0 × 10³ cells/cm².
For determination of the effect of cisplatin on TNBC cells, cells were cultured with or without 10, 20, 50, 100, and 200 μM DDP for 24 h or with 10 μM DDP for 24 h and then collected for subsequent experiments.

Ma H.Y., Li Y., Yin H.Z., Yin H., Qu Y.Y, & Xu Q.Y. (2020). TNFAIP8 Promotes Cisplatin Chemoresistance in Triple-Negative Breast Cancer by Repressing p53-Mediated miR-205-5p Expression. Molecular Therapy. Nucleic Acids, 22, 640-656.

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Breast Cancer Cell Lines Culture

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Breast cancer cell lines (MCF7 and ZR-75 – both ER and MGMT positive, ZR-75-30, HCC 1428 and MDA MB 468 – MGMT positive and ER weakly positive/negative) purchased from the American Tissue Culture Collection (ATCC; Manassas, VA) were grown in DMEM (MCF-7, T-47D, MDA MB 468) and RPMI 1640 (ZR-75-1, ZR-75-30, HCC 1428) medium. All the media were supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin (GIBCO, Invitrogen Corporation, NY). Adherent monolayer cultures were maintained at 37°C containing 5% CO₂.

Bobustuc G.C., Kassam A.B., Rovin R.A., Jeudy S., Smith J.S., Isley B., Singh M., Paranjpe A., Srivenugopal K.S, & Konduri S.D. (2018). MGMT inhibition in ER positive breast cancer leads to CDC2, TOP2A, AURKB, CDC20, KIF20A, Cyclin A2, Cyclin B2, Cyclin D1, ERα and Survivin inhibition and enhances response to temozolomide. Oncotarget, 9(51), 29727-29742.

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Murine Model for Triple Negative Breast Cancer

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4T1 cells (American Type Culture Collection—ATCC), a widely used triple negative breast cancer cell line [18 (link),19 (link)] and a murine model for stage IV human breast cancer, were grown at 37°C with 5% CO₂ in RPMI-1640 supplemented with 10% fetal bovine serum (FBS), 100 units/mL penicillin and 50 g/mL streptomycin, all from GIBCO (Grand Island, NY). MDA-MB-231 and MDA-MB-468, also triple negative breast cancer cell lines, however, derived from human breast tumors, were purchased from ATCC and cultivated at 37°C with 5% CO₂ in DMEM medium supplemented with 10% FBS, 100 units/mL penicillin and 50 g/mL streptomycin. HEK293 cells, used for adenoviral expansion, were grown at 37°C with 5% CO₂ in DMEM supplemented with 10% FBS, 100 units/mL penicillin and 50 g/mL streptomycin, all from GIBCO (Grand Island, NY). Female Balb/C mice at 7–8 weeks (approximately 25-30g) were used for in vivo experiments. Animal care and experimental procedures were complied with Universidade Federal de Minas Gerais—Comissão de Ética no Uso de Animais (CEUA)—institutional animal welfare established guidelines (CEUA/UFMG/Approval 88/2013). Animals were maintained on a standard diet and water access ad libitum and housed under a 12-hour light–dark cycle.

Guimarães E., Machado R., Fonseca M.D., França A., Carvalho C., Araújo e Silva A.C., Almeida B., Cassini P., Hissa B., Drumond L., Gonçalves C., Fernandes G., De Brot M., Moraes M., Barcelos L., Ortega J.M., Oliveira A, & Leite M.F. (2017). Inositol 1, 4, 5-trisphosphate-dependent nuclear calcium signals regulate angiogenesis and cell motility in triple negative breast cancer. PLoS ONE, 12(4), e0175041.

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