Human fibrosarcoma HT1080 cells (ATCC) were cultured in Dulbecco's modified Eagle's medium (DMEM, Mediatech) supplemented with 10% (v/v) fetal bovine serum (FBS, Hyclone Laboratories), and 0.005% (w/v) gentamicin (Quality Biological). Human breast carcinoma MDA-MB-231 cells (ATCC) and MCF-7 cells (ATCC) were cultured in DMEM (Mediatech) supplemented with 10% FBS (Hyclone). Human glioblastoma U-87 MG cells (ATCC) were cultured in DMEM (Mediatech) supplemented with 10% FBS (Hyclone). Human diploid cell line, WI-38, (ATCC) were cultured in Eagle's minimal essential medium (EMEM, Mediatech) supplemented with 10% FBS (Hyclone). Human breast epithelial MCF10A cells (ATCC) and MCF12A cells (ATCC) were cultured in DMEM supplemented with 5% horse serum (Atlanta biologicals), 20 ng ml−1 Human epidermal growth factor (Sigma-Aldrich), 100 ng ml−1 cholera toxin, (Sigma-Aldrich) 0.01 mg ml−1 bovine insulin (Life technologies), and 500 ng ml−1 hydrocortisone (Sigma-Aldrich). HT1080 cells transfected with shRNAs (see below) were grown in medium containing 1 μg ml−1 puromycin. The cells were maintained at 37 °C and 5% CO2 in a humidified incubator during cell culture and during live-cell microscopy. All cell lines were tested for mycoplasma and deemed free of contamination.
Mcf 12a cells
Manufactured by American Type Culture Collection
Sourced in United States
MCF-12A cells are a non-transformed, immortalized human breast epithelial cell line derived from normal human mammary gland tissue. They are commonly used as a model system for studying normal breast epithelial cell biology and physiology.
Automatically generated - may contain errors
Lab products found in correlation
Mcf 7 cell, by American Type Culture Collection (2 mentions)
Mda mb 231 cell, by American Type Culture Collection (2 mentions)
Hydrocortisone, by Merck Group (2 mentions)
Cholera toxin, by Merck Group (2 mentions)
Insulin, by Merck Group (2 mentions)
Bovine insulin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Human epidermal growth factor (hegf), by Merck Group (1 mentions)
Horse serum, by Atlanta Biologicals (1 mentions)
Dulbecco modified eagle medium (dmem), by Atlanta Biologicals (1 mentions)
U87mg cells, by American Type Culture Collection (1 mentions)
Wi 38, by American Type Culture Collection (1 mentions)
Ht1080 cells, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Cytiva (1 mentions)
Gentamicin, by Quality Biological (1 mentions)
Htb 26, by American Type Culture Collection (1 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
5 protocols using mcf 12a cells
1
Cell Culture Conditions for Various Cell Lines
2
MDA-MB-231 and MCF-12A Cell Culture Protocol
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MDA-MB-231 cells (human Caucasian triple negative (ER−, PR−, Her2−), claudin-low breast carcinoma, ATCC HTB-26, from ATCC (Manassas, VA, USA)) were cultured in DMEM-HG cell growth medium supplemented with 10% (v/v) FBS. MCF-12A cells (non-neoplastic breast cell line, ATCC CRL-10782, from ATCC (Manassas, VA, USA)) were cultured in DMEM/F12 medium supplemented with 20 ng/mL hEGF, 100 ng/mL cholera toxin, 0.01 mg/mL bovine insulin, 500 ng/mL hydrocortisone and 5% (v/v) horse serum. Cells were cultured in monolayers in a sterile environment at 37 °C with 5% CO2 humidified atmosphere. Under these conditions the population doubling time was 25.5 ± 0.9 h and 20.6 ± 3.1 h for MDA-MB-231 and MCF-12A cells, respectively. The cell cultures were routinely screened for mycoplasma contamination presenting negative results. Tested compounds were added during the logarithmic phase of cell growth.
3
Breast Cancer Cell Lines: Cultivation and Validation
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Human breast epithelial non-tumorigenic MCF-12A cells and tumorigenic MDA-MB-231, MDA-MB-468, and MDA-MB-453 cells were obtained from ATCC. Authentication of cell lines was performed by short tandem repeat (STR) profiling at Northwestern NUseq Core Facility. The resulting autosomal STR profiles were compared and matched 100% to the ATCC database.
MCF-12A was cultured in Dulbecco’s modified Eagle’s medium (DMEM), and supplemented DMEM/F12 medium (GIBCO) was added with 5% horse serum (GIBCO), 0.02 μg/mL human epidermal growth factor (EGF, Sigma-Aldrich), 0.5 μg/mL hydrocortisone (Sigma-Aldrich), 0.1 μg/mL cholera toxin (Sigma-Aldrich), 10 μg/mL insulin (Sigma-Aldrich), and 1% penicillin-streptomycin (GIBCO). MDA-MB-231, MDA-MB-468, and MDA-MB-453 were cultured in Dulbecco’s modified Eagle’s medium (DMEM) and supplemented with 10% Fetal Bovine Serum (FBS, GIBCO) and 1% penicillin-streptomycin (GIBCO). Cells were grown at 37 °C in a humidified atmosphere of 5% of CO2.
MCF-12A was cultured in Dulbecco’s modified Eagle’s medium (DMEM), and supplemented DMEM/F12 medium (GIBCO) was added with 5% horse serum (GIBCO), 0.02 μg/mL human epidermal growth factor (EGF, Sigma-Aldrich), 0.5 μg/mL hydrocortisone (Sigma-Aldrich), 0.1 μg/mL cholera toxin (Sigma-Aldrich), 10 μg/mL insulin (Sigma-Aldrich), and 1% penicillin-streptomycin (GIBCO). MDA-MB-231, MDA-MB-468, and MDA-MB-453 were cultured in Dulbecco’s modified Eagle’s medium (DMEM) and supplemented with 10% Fetal Bovine Serum (FBS, GIBCO) and 1% penicillin-streptomycin (GIBCO). Cells were grown at 37 °C in a humidified atmosphere of 5% of CO2.
4
MCF-12A cell culture protocol
5
Cell Culture and DNMT Inhibitor Treatment
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Cell culture and treatments. MCF7 cells (ATCC, Maryland USA) were cultured in Eagle's Minimum Essential Medium (Gibco, ThermoFisher Scientific, UK) supplemented with 10% (v/v) foetal bovine serum (Gibco) and 0.01mg/ml insulin (Sigma-Aldrich, Missouri, USA). Hec50 cells (ATCC, Maryland USA) were cultured in Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 (DMEM/F12) (Gibco, ThermoFisher Scientific, UK) supplemented with 10% (v/v) fetal bovine serum (Gibco), 1% (v/v) penicillin and streptomycin (Gibco), sodium bicarbonate and sodium pyruvate. MCF12A cells (ATCC, Maryland USA) were cultured in Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 (DMEM/F12) (Gibco, ThermoFisher Scientific, UK) supplemented with 5% (v/v) horse serum (Gibco), 20ng/ml human epidermal growth factor, 100 ng/ml cholera toxin, 0.01 mg/ml bovine insulin and 500 ng/ml hydrocortisone 95%. All cell lines were grown at 37°C in a humidified atmosphere with 5% CO2 to 90% confluency in T75 flasks (Corning, New York, USA) before collection. Cells were grown to 40% or 60% confluency respectively before exchanging full growth media for stripped media prior to treatment with the DNMT inhibitor 5-Aza-2′-deoxycytidine (1 µM), suberoylanilide hydroxamic acid (2.5 µM) or vehicle (DMSO).
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