The normal epithelial breast cell line, MCF-10A, was obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA) and grown in EMBM (ATCC) with 10% fetal bovine serum (FBS; Gibco, Carlsbad, CA, USA) and 1% penicillin/streptomycin (Gibco), plus 100 ng/ml cholera toxin (ATCC). Two human breast cancer cell lines, BT474 and MDA-MB-231, were obtained from Cell Bank of the Chinese Academy of Sciences (Shanghai, P.R. China) and grown in RPMI-1640 medium (Hyclone, Logan, UT, USA) with 10% FBS and antibiotics. All cells were cultivated at 37°C in a 5% CO2 incubator.
Cholera toxin
Manufactured by American Type Culture Collection
Sourced in United States
Cholera toxin is a protein produced by the bacterium Vibrio cholerae, the causative agent of cholera. It functions as an exotoxin, which is a toxin secreted by bacteria that can affect the host organism. The core function of cholera toxin is to stimulate the release of water and electrolytes from intestinal cells, leading to the severe diarrhea characteristic of cholera.
Automatically generated - may contain errors
Lab products found in correlation
Insulin, by Merck Group (4 mentions)
Hydrocortisone, by Merck Group (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Cholera toxin, by Merck Group (3 mentions)
Mcf 10a, by American Type Culture Collection (2 mentions)
Penicillin streptomycin, by GE Healthcare (2 mentions)
Dmem basal medium, by Thermo Fisher Scientific (2 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (2 mentions)
Horse serum, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by GE Healthcare (2 mentions)
Dmem f12 basal medium, by Thermo Fisher Scientific (2 mentions)
Bt 474, by National Collection of Authenticated Cell Cultures (1 mentions)
Mda mb 231, by National Collection of Authenticated Cell Cultures (1 mentions)
Skbr3, by American Type Culture Collection (1 mentions)
Bt549, by American Type Culture Collection (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
Mda mb 468, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Biowest (1 mentions)
Sk br 3, by Biowest (1 mentions)
6 protocols using cholera toxin
1
Culturing Normal and Breast Cancer Cells
2
Mammary Epithelial Cell Culture Protocols
3
Culturing Diverse Breast Cell Lines
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The human mammary epithelial cell line MCF10A and human breast cancer cell lines MCF7, T-47D, SK-BR-3, MDA-MB-231, MDA-MB-468 and BT-549 were bought from ATCC (Rockville, MD, USA). MCF10A was cultured in MEGM with100 ng/ml cholera toxin (ATCC, USA). MCF7, SK-BR-3 and MDA-MB-468 were cultured in DMEM with 10% fetal bovine serum (FBS, Biowest). T-47D and BT-549 were cultured in RPMI 1640 with 10% FBS. MDA-MB-231 was cultured in L-15 with 10% FBS. All cell lines were cultured at 37 °C in a humidified atmosphere containing 5% CO2.
4
Cultivation and Maintenance of Colon Cell Lines
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All human cell lines were purchased from American Type Culture Collection (ATCC). The normal colon epithelial cell line FHC was cultured in DMEM/F12 medium (cat. no. 30-2006; ATCC) with a final concentration of 25 mM HEPES, 10 ng/ml cholera toxin (cat. no. C8052; Sigma-Aldrich; Merck KGaA), 0.005 mg/ml insulin (cat. no. 91077C; Sigma-Aldrich; Merck KGaA), 0.005 mg/ml transferrin (cat. no. T8158; Sigma-Aldrich; Merck KGaA), 100 ng/ml hydrocortisone (cat. no. HY-N0583; MedChemExpress), 20 ng/ml human recombinant EGF (cat. no. PHG0311; Thermo Fisher Scientific, Inc.) and 10% (v/v) FBS (cat. no. 10099141C; Thermo Fisher Scientific, Inc.). HCT-8 and HCT-116 cells were cultured in RPMI-1640 medium (Thermo Fisher Scientific, Inc.) supplemented with 10% (v/v) FBS; SW620 cells were cultured in Leibovitz's L-15 medium (Thermo Fisher Scientific, Inc.) supplemented with 10% (v/v) FBS. All cells except SW620 (cultured with no CO2 at 37˚C) were maintained in a humidified atmosphere with 5% CO2 at 37˚C. All treated cells were incubated at 37˚C in the following experiments.
5
Culturing Breast Cancer Cell Lines
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Human breast cancer cell lines T-47D (HTB-113; ATCC), Hs 578T (HTB-126; ATCC) and MDA-MB-231 (HTB-26; ATCC) were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. MCF-10-2A (CRL-10781; ATCC) cells were grown in DMEM-F12 medium supplemented with 5% horse serum, 1% penicillin/streptomycin, 20 ng/ml epidermal growth factor (EGF), 100 ng/ml cholera toxin, 0.01 mg/ml insulin and 500 ng/ml hydrocortisone. All cell lines were maintained at 37°C in a humidified atmosphere with 5% CO2. For all experiments, cells were seeded in 6-well at a concentration of 1.5×105 cells/ml for 24 h experiments and 1×105 cells/ml for 48 h experiments, with the exception of the ECAR experiments in which cells were seeded in XF 24-well plates at a concentration of 1×104 cells/well. All medium constituents were acquired from Biochrom with the exception of EGF, cholera toxin, hydrocortisone and insulin that were purchased from Sigma-Aldrich.
6
Mammary Epithelial Cell Culture Protocols
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MCF10A mammary epithelial cells were obtained from ATCC and cultured according to established protocols48 (link). DMEM/F12 basal medium (ThermoFisher Scientific) was supplemented with 5% horse serum (ThermoFisher Scientific), 1% penicillin/streptomycin (ThermoFisher), 20 ng/ml epidermal growth factor (Peprotech), 0.5 mg/ml hydrocortisone (Sigma), 100 ng/ml cholera toxin (Sigma), and 10 μg/ml insulin (Sigma). HME1 cells were obtained from ATCC and cultured in the medium described above without cholera toxin. MCF7 and MDA-MB-231 cell lines were obtained from ATCC and cultured in DMEM basal medium (ThermoFisher Scientific) supplemented with 10% fetal bovine serum (GE Healthcare), and 1% penicillin/streptomycin. Cells were encapsulated at 50,000 cells/ml final concentration in hydrogel matrices and cultured for 14 days.
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