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Mda mb 231 cells

Manufactured by PerkinElmer
Sourced in United Kingdom, United States

MDA-MB-231 cells are a human breast cancer cell line commonly used in research. They are triple-negative breast cancer cells, meaning they lack expression of estrogen receptor, progesterone receptor, and HER2. MDA-MB-231 cells are frequently utilized for various in vitro and in vivo studies related to breast cancer biology, including cancer cell proliferation, migration, invasion, and drug screening.

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3 protocols using mda mb 231 cells

1

Cell Line Procurement and Cultivation

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Authenticated MDA-MB-231, MDA-MB-157 and PC3 lines were obtained from ATCC (Teddington, UK). Hs578T cells were kindly provided by Dr Paul Mullan (Centre for Cancer Research and Cell Biology, Queen's University Belfast). The highly invasive clone, MDA-MB-231BO was provided complete with matched parental cells by Prof. Toshiyuki Yoneda (University of Osaka, Japan) [30 (link)] and were cultured in DMEM supplemented with 10% v/v foetal calf serum (FCS) (Invitrogen™ Life Technologies, Paisley, UK). CD44 depleted luciferase-expressing MDA-MB-231 cells from Caliper Life Sciences (Cheshire, UK) were generated in our laboratory by stable transfection with CD44 shRNA or non-targeting shRNA control as previously described [7 (link)] and cultured in DMEM supplemented with 10% v/v FCS, 0.2 μg/ml puromycin. All cell lines were grown to 70% confluence prior to experimentation. Human bone marrow endothelial cells (BMEC) were provided by Dr Babette Weksler (Cornell University, NY) and cultured as previously described [10 (link)].
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2

Cell Line Maintenance and Characterization

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All cell lines were maintained at 37°C and 5% CO2, and passaged at 95-100% confluence. The following cell lines were used in this study: MDA-MB-231-luc D3 H2LN, B16-F1 and HEK-293T. B16-F1 and HEK-293T were originally obtained from the ATCC, while MDA-MB-231 cells were obtained from Caliper Life Sciences. MDA-MB-231 and B16-F1 cells were grown in MEM-alpha media, 10% FBS (Gibco) and 1% pen/strep. HEK-293T cells were grown in low glucose, pyruvate DMEM, 10% FBS (Gibco) and 1% pen/strep. All cell lines were tested to be Mycoplasma-negative using the MycoAlert™ Mycoplasma detection kit (Lonza).
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3

Culturing Human Breast Cancer and Non-Malignant Cells

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Human breast cancer and non-malignant epithelial cells were obtained from the American Type Culture Collection (Rockville, MD, USA). MDA-MB-231 cells stably expressing luciferase (MDA-MB-231-Luc) were obtained from Caliper Lifesciences (Hopkinton, MA, USA). All cell culture media contained 10% FBS and 1% penicillin–streptomycin unless otherwise specified. Human MCF-7, MDA-MB-231 and MDA-MB-435 cells were grown in DMEM media. The MCF-7 p53 KD and MCF-7 MDM2 KD inducible cells were generated using the previously described method53 (link),54 (link) and were grown in the same media as MCF-7 cells, but supplemented with 0.5 μg ml −1 puromycin or 10 μg ml −1 blasticidin and 0.5 μg ml −1 puromycin (Sigma-Aldrich Co.), respectively. MDAMB-468 cells were grown in DMEM/F-12 Ham’s media (DMEM/F-12 1:1 mixture). The MCF-10A growth medium was composed of DMEM/F-12, supplemented with 5% donor horse serum, 20 ng ml −1 epidermal growth factor, 10 μg ml −1 insulin, 0.5 μg ml −1 hydrocortisone (Sigma-Aldrich Co.) and 100 μg ml −1 cholera toxin (Cambrex, Westborough, MA, USA). Human MDAMB-231-Luc cells were grown in DMEM media containing 0.1 mM MEM non-essential amino acids and 2 mM L-glutamine.
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