The MCF-7 human breast adenocarcinoma cell line was procured via the American Type Culture Collection (ATCC, Manassas, VA, USA). Roswell Park Memorial Institute medium (RPMI-1640) supplemented with 10% fetal bovine serum and 1% penicillin and streptomycin (Sigma-Aldrich, St. Louis, MO, USA) was used for the cultivation of MCF-7 and this were subsequently cultured in a humidified incubator using 5% CO2 at 37 °C.
Mcf 7 human breast adenocarcinoma cell line
Manufactured by American Type Culture Collection
Sourced in United States
The MCF-7 human breast adenocarcinoma cell line is a well-established in vitro model derived from a pleural effusion of a 69-year-old Caucasian woman with metastatic breast adenocarcinoma. This cell line exhibits epithelial-like morphology and is commonly used in breast cancer research.
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Lab products found in correlation
Rpmi 1640, by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Mcf 7, by GeneCopoeia (1 mentions)
Dmem medium, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Sk ov 3 human ovarian adenocarcinoma cell line, by American Type Culture Collection (1 mentions)
Hela human cervical adenocarcinoma cell line, by American Type Culture Collection (1 mentions)
Human mda mb 231 breast adenocarcinoma cell line, by American Type Culture Collection (1 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Mcf 10a, by American Type Culture Collection (1 mentions)
6 protocols using mcf 7 human breast adenocarcinoma cell line
1
MCF-7 Human Breast Cancer Cell Culture
2
Modulating LGR5 in Breast Cancer Cells
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The MCF-7 human breast adenocarcinoma cell line was purchased from the American Type Culture Collection (Manassas, VA). The MDA-MB-453 human breast cancer cell line was purchased from Thermo Fisher Scientific (Shanghai, China). MCF-7 and MDA-MB-453 cells were cultured in DMEM medium supplemented with 10% FBS, 100 U/ml penicillin, and 0.1 g/ml streptomycin, in a cell incubator with humidified atmosphere (37%, 5% CO2). DMEM medium, FBS, and penicillin-streptomycin for cell culture were purchased from Sigma (Merck Life Science, Shanghai, China). LGR5 overexpression in MCF-7 cells as well as LGR5 knockdown in MDA-MB-453 cells were performed by Genecopoeia (Guangzhou, China). Briefly, for LGR5 overexpression, LGR5 cDNA-carrying pcDNA3.1 vector was transfected into MCF-7 cells, and cells transfected with empty vector were used as wild-type control (WT). For LGR5 knockdown, short hairpin RNA (shRNA) targeting LGR5 mRNA was transfected into MDA-MB-453 cells, and cells transfected with non-targeting shRNA was used as wild-type control (WT). Equal LGR5 mRNA and protein expression in non-transfected cells and transfected control cells were confirmed by RT-qPCR and Western blot (data not shown).
3
Cytotoxicity of Blank and Gem-Loaded Magnetic PCL Particles
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Cytotoxicity of blank (Gem unloaded) magnetic PCL particles was investigated in the CCD-18 human colon fibroblast cell line (Scientific Instrumentation Centre, University of Granada, Granada, Spain), and in the MCF-7 human breast adenocarcinoma cell line (American Type Culture Collection, ATCC, Manassas, VA, USA). Triplicate cultures were evaluated by the 3-(4,5-dimethylthiazol-2-yl)-3,5-diphenyl tetrazolium bromide (MTT) proliferation assay. Detailed description of the procedure can be found in the literature [39 (link)]. Cell lines were kept in contact with NP concentrations, ranging from 0.05 to 100 μg/mL, for 48 and 72 h.
The MTT assay was also used to define the cytotoxicity against MCF-7 cells of the Gem-loaded Fe3O4/PCL particles (of 2:4 weight ratio) in comparison with free Gem (n = 3). The concentration of these nanoformulations in contact with cells ranged from 0.1 to 50 µM equivalent Gem concentrations.
Cells without treatment were used as a control to calculate the relative cell viability (%) (Equation (4)).
The MTT assay was also used to define the cytotoxicity against MCF-7 cells of the Gem-loaded Fe3O4/PCL particles (of 2:4 weight ratio) in comparison with free Gem (n = 3). The concentration of these nanoformulations in contact with cells ranged from 0.1 to 50 µM equivalent Gem concentrations.
Cells without treatment were used as a control to calculate the relative cell viability (%) (Equation (4)).
4
MCF7 Breast Cancer Cell Culture
5
Intracellular ROS Detection by HSN in Cell Lines
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4T1 murine mammary carcinoma cell line, MCF-7 human breast adenocarcinoma cell line, HepG2 human hepatocellular carcinoma cell line, NIH/3T3 murine fibroblast cell line, NDF normal human dermal fibroblast cell line, MDA-MB-231 human breast adenocarcinoma cell line, PC12 rat pheochromocytoma cell line, HeLa human cervical adenocarcinoma cell line, and SKOV3 human ovarian adenocarcinoma cell line (purchased from American Type Culture Collection, ATCC) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% antibiotics (penicillin and streptomycin) and placed in a humid CO2 incubator providing an atmosphere containing 5% CO2 at 37 °C.
To detect the intracellular ROS aroused by HSN, cells were seeded in confocal cell culture dishes at a density of 1 × 104 cells per dish. After culture in incubator for 12 h, HSN (final concentration: [pTBCB] = 50 µg mL−1) or PBS were added to the cells and incubated with cells for 24 h. Thereafter, cells were gently washed three times with fresh PBS to remove excess nanoparticles, and then fixed with 4% paraformaldehyde. After fixation, cells were successively stained with DAPI and DCF-DA. Then cells were imaged under a LSM800 confocal microscope.
To detect the intracellular ROS aroused by HSN, cells were seeded in confocal cell culture dishes at a density of 1 × 104 cells per dish. After culture in incubator for 12 h, HSN (final concentration: [pTBCB] = 50 µg mL−1) or PBS were added to the cells and incubated with cells for 24 h. Thereafter, cells were gently washed three times with fresh PBS to remove excess nanoparticles, and then fixed with 4% paraformaldehyde. After fixation, cells were successively stained with DAPI and DCF-DA. Then cells were imaged under a LSM800 confocal microscope.
6
Culturing MCF-7 and MCF-10A Cell Lines
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The MCF-7 human breast adenocarcinoma cell line was purchased from the American Type Culture Collection (ATCC) (LGC standards, Middlesex, UK). Cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% (v/v) foetal bovine serum, 2 mM glutamine, 50 U/ml penicillin and 50 U/ml streptomycin and 1% non-essential amino acids. The MCF-10A, non-tumorigenic breast epithelial cell line was purchased from ATCC (LGC standards, Middlesex, UK). Cells were maintained in Ham’s F12:DMEM (1:1), 20 ng/ml epidermal growth factor (EGF) (PeproTech, London, UK), 0.1 µg/ml cholera toxin, 10 µg/ml insulin, 500 ng/ml hydrocortisone, 5% horse serum and 50 U/ml penicillin and 50 U/ml streptomycin.
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