SK-BR3, SK-OV3, BT-474, MCF7, MDA231, MDA468, HCC2281, MDA-361, MDA-453, HCC-1419, HCC-1569, UACC-812, LnCap, MDA-175, MCF-10A, HCC38 and HG261 cell lines were purchased from American Type Culture Collection and cultured as instructed. Primary cell lines (keratinocytes, osteoblast, renal epithelial, pulmonary artery endothelial cells, pulmonary artery smooth muscle, neural progenitor, CD34+ enriched PBMC) were obtained from Promocell and cultured according to their protocols. Primary lymphocytes were isolated from normal donors provided by the University of Pennsylvania Human Immunology Core and cultured in R10 medium (RPMI 1640 supplemented with 10% fetal calf serum; Invitrogen). Primary lymphocytes were stimulated with microbeads coated with CD3 and CD28 stimulatory antibodies (Life Technologies, Grand Island, NY, Catalog) as described (15 (link)). T cells were cryopreserved at day 10 in a solution of 90% fetal calf serum and 10% dimethylsulfoxide (DMSO) at 1 × 108 cells/vial.
Mda 453
Manufactured by American Type Culture Collection
Sourced in United States, Canada
The MDA-453 is a cell line derived from a human breast carcinoma. It is a commonly used model for the study of breast cancer biology and drug development.
Automatically generated - may contain errors
Lab products found in correlation
Mcf 7, by American Type Culture Collection (5 mentions)
Skbr3, by American Type Culture Collection (4 mentions)
Mda 231, by American Type Culture Collection (3 mentions)
Bt474, by American Type Culture Collection (2 mentions)
Skov3, by American Type Culture Collection (2 mentions)
Mda 468, by American Type Culture Collection (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Lncap, by American Type Culture Collection (1 mentions)
Mcf 10a, by American Type Culture Collection (1 mentions)
Hcc38, by American Type Culture Collection (1 mentions)
Hcc1419, by American Type Culture Collection (1 mentions)
Uacc 812, by American Type Culture Collection (1 mentions)
Hcc1569, by American Type Culture Collection (1 mentions)
Mda 361, by American Type Culture Collection (1 mentions)
Dulbecco s modified eagle s medium, by Merck Group (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
Dms 273, by American Type Culture Collection (1 mentions)
Fetal calf serum (fcs), by Biowest (1 mentions)
Hbmec, by Lonza (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
7 protocols using mda 453
1
Culturing Cell Lines and Primary Cells
2
Establishment and Maintenance of Cell Lines
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Human cancer cell lines SKBR3, PM1, MDA-453, MDA-468, DMS-273, PC3, FM-88, and embryonic kidney HEK293T were purchased from the American Type Culture Collection. Cells were cultured in Roswell Park Memorial Institute medium 1640 (Sigma St Louis, MO) or Dulbecco’s Modified Eagle’s medium (Sigma St Louis, MO) supplemented with 10% heat-inactivated fetal calf serum and antibiotics. Normal human mammary epithelial cells and normal lung fibroblasts were purchased from Clonetics/BioWittaker (Allendale, NJ) and maintained in complete Mammary Epithelial Cell Growth medium (Lonza, Verviers, Belgium). Human peripheral blood mononuclear cells were prepared from buffy coats obtained from normal volunteers. Monocytes were enriched from peripheral blood mononuclear cells using plastic adherence, and purification was verified by phenotypic analysis of the surface marker CD14+. Bone marrow–derived mesenchymal stem cells were prepared from bone marrow aspirates as described previously.51 (link) Approval for obtaining blood and bone marrow samples from normal volunteers was granted by Oslo University Hospital Ethics Committee.
3
Characterization of Prostate and Breast Cancer Cell Lines
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Experiments were carried on canonical human prostate cancer cell line PC-3 (American Type Culture Collection, Rockville, MD, USA) Cells were cultured in RPMI supplemented with 10% FBS, 1% penicillin and 0.16% kanamycin and grown in a humidified 5% CO2 incubator at 37 °C. Cells were regularly authenticated in terms of cell viability, morphology and doubling time. For Her2 characterization, breast cancer cell lines, i.e., MDA231, MDA 453 and SKBR3, were used (American Type Culture Collection, Manassas, VA, USA).
4
Cell Line Cultivation for Breast Cancer
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Breast cancer cell lines including MCF-7, MDA-231, T47D and MDA-453 were originally obtained from American Type Culture Collection (Manassas, VA); Doxorubicin-resistant MCF-7 (MCF-7R) cells [16 (link)] were provided by Dr. Zhiyuan Zhong, Soochow University; HEK293 cells were obtained from Dr. Aaron Schimmer (the University of Toronto, Canada). Breast cancer cells and HEK293 cells were maintained in RPMI-1640 and Dulbecco's high glucose modified Eagle's medium (DMEM) (Hyclone), respectively. All media were supplemented with 10% fetal calf serum (Biowest®, Nuaillé, France), 100 μg/ml penicillin, and 100 U/mL streptomycin.
5
Breast and Ovarian Cancer Cell Line Treatment
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MCF‐7, BT474, MDA‐453, and SKOV3 cells were obtained from the American Type Culture Collection. MCF‐7/Her2 cells stably overexpressing Her2 were established in our laboratory as described previously.28 For treatment with β‐AR agonists and antagonist, the cells were incubated overnight in serum‐free medium and then treated with 10 μmol/L epinephrine, 10 μmol/L norepinephrine, 5 or 10 μmol/L isoproterenol (ISO), 0.5 or 2 μmol/L ICI‐118551, 1 or 5 μmol/L DAPT or 1 μmol/L Atenolol (ATEN) for the indicated time points. For treatment with γ‐secretase antagonist, the cells were treated with 0.5 or 2 μmol/L L685458.
6
Establishing and Characterizing Breast Cancer Metastatic Cell Lines
7
Breast Cancer Cell Line Proliferation Assay
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Breast cancer cell lines MCF7, ZR75, MDA435, MDA453, MB231, BT20, HS578T, and HCC1937 were purchased from American Type Culture Collection (ATCC) and maintained in standard conditions. Transfection was performed with Lipofectamine 2000 (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. Targeted sequences for small interfering RNA (siRNA)-induced silencing were all listed in Supplementary Table 2 .
Cell suspension (100 μL/well) was inoculated in a 96-wellplate, pre-incubated in a 37°C humidified incubator (5% CO2). After each of the 0, 24, 48, and 72 h time points, 10 μL of the CCK8 reagent from Sigma (St.Louis, MO) was added to each well of the corresponding plate. The plate was incubated for two additional hours and the 450nm absorbance was measured.
Cell suspension (100 μL/well) was inoculated in a 96-wellplate, pre-incubated in a 37°C humidified incubator (5% CO2). After each of the 0, 24, 48, and 72 h time points, 10 μL of the CCK8 reagent from Sigma (St.Louis, MO) was added to each well of the corresponding plate. The plate was incubated for two additional hours and the 450nm absorbance was measured.
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