DMEM and RPMI 1640 cell culture media, antibiotics, and nonessential amino acids were purchased from Life Technologies (Grand Island, NY). CD14+ monocytes were isolated by MACS CD14 microbeads from Miltenyi Biotec (Auburn, CA). Human Abs, including CD16 allophycocyanin, CD16 FITC, CD14 FITC, CD40 FITC, and CD86 FITC, were purchased from eBioscience (San Diego, CA). Abs CD14 allophycocyanin, CD14 PE, CD163 PE, CD11c allophycocyanin, CD68 PE, CD206 allophycocyanin, DC-SIGN FITC, and isotype control Abs were purchased from BD Pharmingen (Franklin Lakes, NJ). Phospho-p44/42-ERK1/2 and anti-mouse IgG PE were obtained from Cell Signaling Technology (Danvers, MA). Human IL-10 Ab and mouse IgG1 isotype control were from R&D Systems (Minneapolis, MN). miR-27a inhibitor, mimic, and scrambled controls were purchased from Ambion Life Technologies (Carlsbad, CA). Lipofectamine RNAiMAX transfection reagent was from Life Technologies. The sprouty2 construct was obtained from OriGene (Rockville, MD), which was transfected by Roche (Indianapolis, IN) X-tremeGENE transfection reagent. ERK inhibitor, U0126, was procured from EMD Millipore (Billerica, MA).
Dmem and rpmi 1640 cell culture media
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom
DMEM and RPMI 1640 are commonly used cell culture media formulations. They provide nutrients and growth factors essential for the maintenance and propagation of various cell types in vitro. The core function of these media is to support the growth and proliferation of cells in a controlled laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Cd16 fitc, by Thermo Fisher Scientific (1 mentions)
U0126, by Merck Group (1 mentions)
Cd86 fitc, by Thermo Fisher Scientific (1 mentions)
Cd14 fitc, by Thermo Fisher Scientific (1 mentions)
Cd163 pe, by BD (1 mentions)
Cd14 pe, by BD (1 mentions)
Cd40 fitc, by Thermo Fisher Scientific (1 mentions)
Cd14 allophycocyanin, by BD (1 mentions)
Cd68 pe, by BD (1 mentions)
X tremegene transfection reagent, by Roche (1 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (1 mentions)
Macs cd14 microbeads, by Miltenyi Biotec (1 mentions)
Cd11c allophycocyanin, by BD (1 mentions)
Dc sign fitc, by BD (1 mentions)
T24 bladder cancer cells, by American Type Culture Collection (1 mentions)
Glutamine, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (1 mentions)
Mccoy s 5a modified medium, by American Type Culture Collection (1 mentions)
Gapdh, by Abcam (1 mentions)
4 protocols using dmem and rpmi 1640 cell culture media
1
Monocyte Isolation and Characterization
2
Prostate and Bladder Cancer Cell Culture
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VCaP, LNCaP, and NCI‐H660 prostate cancer cells, T24 bladder cancer cells, McCoy's 5 A Modified Medium, and fetal bovine serum (FBS) were purchased from ATCC (Manassas, VA). DMEM and RPMI‐1640 cell culture media and glutamine were purchased from Life technologies (Carlsband, CA). VCaP cells were maintained in DMEM supplemented with 10% FBS. LNCaP cells were maintained in RPMI‐1640 supplemented with 10% FBS, 2.8 mM l ‐glutamine. NCI‐H660 cells were maintained in RPMI‐1640 medium supplemented with 5% FBS, 4 mM l ‐glutamine, 0.005 mg/ml insulin, 0.01 mg/ml transferrin, 30 nM sodium selenite, 10 nM β‐estradiol, and 10 nM hydrocortisone. T24 bladder cancer cells were maintained in McCoy's 5 A Modified Medium, supplemented with 5% FBS. All cells were cultured in a 5% CO2 humidified incubator at 37°C.
3
Esophageal Cancer Cell Apoptosis Assay
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Eca-109 human esophageal cancer cells and HPT-1A normal human esophageal epithelial cells were provided by the Cell Resource Center of the Shanghai Life Sciences Institute, Chinese Academy of Sciences (Shanghai, China). Other materials included CPI-455 (MSDS, 1628208-23-0; Gibco, Ltd., United States), DMEM and RPMI 1640 cell culture media and fetal bovine serum (Gibco, Ltd). Anti-human P53 (cat no. sc-6243), Bax, KDM5C (cat no. sc-81623 ), Caspase-9 (cat no. 9746), Caspase-3 (cat no. 9502), GAPDH (glyceraldehyde-3-phosphate dehydrogenase, cat no. sc-47778) polyclonal antibody (1:200; Abcam, United Kingdom) were used for western blotting. Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis detection kits (cat no. F7250) were from Sigma-Aldrich (United States). polyvinylidene difluoride (PVDF) membranes were from Biyuntian Biotech (China) and bicinchoninic acid (BCA) protein quantitative detection kits were from Thermo Fisher (United States). An Epics Ultra flow cytometer (Beckman Coulter, United States), JEM-100sx transmission electron microscope (Jeol Ltd., Japan), and a TCS SP2 laser confocal microscope (Leica, Germany) were used.
4
Establishing Human Breast Cancer Cell Lines
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The human breast cancer cell lines (BT-474, SK-BR-3, MDA-MB-453, MDA-MB-231, and MCF-7) were obtained from the Iranian Biological Resource Centre (IBRC, Tehran, Iran). DMEM and RPMI-1640 cell culture media, fetal bovine serum (FBS), and penicillin-streptomycin were purchased from Gibco, BRL (Life technology, Paisley, Scotland). Trichloroacetic acids (TCA) and phenylmethylsulfonyl fluoride (PMSF) were obtained from Sigma Chem. Co (Germany). Also, 2,4-Dinitrophenylhydrazine (DCFH-DA) was obtained from a Molecular probe (Eugene, Oregon, USA). Dimethyl sulfoxide (DMSO) and thiobarbituric acid (TBA) were obtained from Merck (Darmstadt, Germany).
Cell culture BT-474, MDA-MB-453, and MDA-MB-231 cells were maintained in RPMI-1640 medium with 10% fetal bovine serum (FBS). SK-BR-3 and MCF-7 cells were maintained in the Dulbecco's minimum Eagle's media (DMEM) with 10% FBS. BT-474 and SKBR3 media were supplemented with 2mM L-Glutamine, as well. Penicillin (100U/ml) and streptomycin (100μg/ml) were used to control bacterial contamination in all cell lines culture media. The cells cultured in the T-25 flask were maintained at 37°C in the humidified atmosphere with 5% CO2 and passaged every two days.
Cell culture BT-474, MDA-MB-453, and MDA-MB-231 cells were maintained in RPMI-1640 medium with 10% fetal bovine serum (FBS). SK-BR-3 and MCF-7 cells were maintained in the Dulbecco's minimum Eagle's media (DMEM) with 10% FBS. BT-474 and SKBR3 media were supplemented with 2mM L-Glutamine, as well. Penicillin (100U/ml) and streptomycin (100μg/ml) were used to control bacterial contamination in all cell lines culture media. The cells cultured in the T-25 flask were maintained at 37°C in the humidified atmosphere with 5% CO2 and passaged every two days.
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