Dulbecco s minimum essential media dmem

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Dulbecco's Minimum Essential Media (DMEM) is a commonly used cell culture medium formulation. It provides a balanced salt solution and essential nutrients to support the growth and maintenance of a variety of cell types in vitro.

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5 protocols using dulbecco s minimum essential media dmem

Establishment and Maintenance of Sarcoma Cell Lines

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Soft tissue sarcoma (STS) cell lines (SK-LMS, SW982, RD, SW872) as well as primary sarcoma cell lines (ssRMS, BR-CS and WW-LMS) isolated from consented rhabdomyosarcoma patients` primary tumors, were a gift of Dr. med. C. Hinterleitner and Prof. G. Kopp (University of Tübingen).
Cell lines SK-LMS, SW982, RD, ssRMS, BR-CS, and WW-LMS were maintained in Dulbecco’s Minimum Essential Media (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich), 1% penicillin–streptomycin (Biochrom), 1% Sodium pyruvate and 1% MEM-Non-Essential Amino acids (100X) (Gibco), while the SW872 was maintained in RPMI supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich), 1% penicillin–streptomycin (Biochrom), 1% Sodium pyruvate and 1% MEM-Non-Essential Amino acids (100X) (Gibco).
HEK239T cells used for lentiviral pseudo-virus production were obtained from ThermoFisher Scientific and maintained in Hyclone-DMEM medium supplemented with 10% FBS and 200 µM L-glutamine.
All cell lines were cultivated at 37 °C in 5% CO2 humidity.

Tsintari V., Walter B., Fend F., Overkamp M., Rothermundt C., Lopez C.D., Schittenhelm M.M, & Kampa-Schittenhelm K.M. (2022). Alternative splicing of Apoptosis Stimulating Protein of TP53-2 (ASPP2) results in an oncogenic isoform promoting migration and therapy resistance in soft tissue sarcoma (STS). BMC Cancer, 22, 725.

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Biochemical Reagents for Cell Experiments

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Glutaraldehyde (GA), fluorescein isothiocyanate (FITC), manganese chloride tetra-hydrate (MnCl₂·4H₂O), sodium carbonate (Na₂CO₃), phosphate buffered saline (PBS) pH 7.4, glycine, and sodium borohydride (NaBH₄) were purchased from Sigma-Aldrich (Munich, Germany). Ethylene diamine tetra-acetic acid (EDTA) was purchased from Fluka (Buchs, Switzerland). Sterile NaCl solution, 0.9% was purchased from Fresenius Kabi Deutschland GmbH (Bad Humburg, Germany). Sodium hydroxide (NaOH) and dimethyl sulfoxide (DMSO) were purchased from Carl Roth GmbH (Karlsruhe, Germany). Human serum albumin solution 20% was purchased from Grifols Deutschland GmbH (Frankfurt, Germany). Sodium bicarbonate (NaHCO₃) was purchased from RCI Labscan (Bangkok, Thailand). Lipopolysaccharide from Escherichia coli O111:B4 (LPS) was purchased from Sigma-Aldrich (Saint Louis, MO, USA). Dulbecco’s Minimum Essential Media (DMEM) and Roswell Park Memorial Institute 1640 Medium (RPMI), penicillin/streptomycin, and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). MTT reagent (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) was purchased from Bio Basic (Markham, ON, Canada).

Jantakee K., Prapan A., Chaiwaree S., Suwannasom N., Kaewprayoon W., Georgieva R., Tragoolpua Y, & Bäumler H. (2021). Fabrication and Characterization of Human Serum Albumin Particles Loaded with Non-Sericin Extract Obtained from Silk Cocoon as a Carrier System for Hydrophobic Substances. Polymers, 13(3), 334.

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Stem Cell Osteogenic and Angiogenic Differentiation

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Methylcellulose with a viscosity of 15 cP (2% w/v in water at 20 °C), sodium alginate, chitosan (75−85% deacetylation), sodium tripolyphosphate (85%), β-glycerol phosphate (≥98.0%), L-ascorbic acid (cell culture tested), and dexamethasone were all purchased from Sigma-Aldrich (St. Louis, MO). Calcium chloride was purchased from Fisher Scientific (Waltham, MA). Recombinant BMP-9 and VEGF along with their corresponding ELISA kits were purchased from R&D Systems (Minneapolis, MN). Human bone marrow mesenchymal stem cells at passage 4, xenofree media (StemPro), Dulbecco’s minimum essential media (DMEM, Gibco), alpha minimum essential media (α-MEM), fetal bovine serum (FBS, Gibco), penicillin−streptomycin (Gibco), gentamicin (Gibco), and live and dead cell assay kit (Invitrogen) were all purchased from ThermoFisher Scientific (Waltham, MA). Alkaline phosphatase (ALP) assay kit was purchased from BioVision Incorporated (San Francisco, CA).

Zhou X., Berglund P., Zhao H., Liljeström P, & Jondal M. (1995). Generation of cytotoxic and humoral immune responses by nonreplicative recombinant Semliki Forest virus.. Proceedings of the National Academy of Sciences of the United States of America, 92(7), 3009-3013.

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Culturing U87 MG Glioblastoma Cell Line

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The human U87 MG glioblastoma cancer (ATCC^® HTB-14™) cell line was obtained from the American Type Culture Collection (ATCC, Virginia, United States). Dulbecco's Minimum Essential Media (DMEM), Eagle’s Minimum Essential Media (EMEM), Fetal bovine serum (FBS) and Phosphate Buffer Saline (PBS) were purchased from Life technologies (Pty) Ltd. (Fairlands, Johannesburg, RSA). The 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT), Dimethyl Sulfoxide (DMSO), Trypsin, and all other chemicals and reagents were of analytical grade and acquired from Merck (Pty) Ltd. (Modderfontein, Johannesburg, RSA). The U87 cell line was maintained in DMEM:EMEM (1:1), supplemented with 10% heat-inactivated FBS and grown at 37°C in a humidified incubator set at 5% CO2. Cells were sub-cultured with 0.25% (w/v) trypsin 0.53 mM ethylenediaminetetraacetic acid (EDTA) for a maximum of 15 min every 2 and 3 days after they had formed an 80% confluent monolayer.

Erukainure O.L., Matsabisa M.G., Salau V.F, & Islam M.S. (2020). Tetrahydrocannabinol-Rich Extracts From Cannabis Sativa L. Improve Glucose Consumption and Modulate Metabolic Complications Linked to Neurodegenerative Diseases in Isolated Rat Brains. Frontiers in Pharmacology, 11, 592981.

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Isolation and Culture of Murine Macrophages and Dendritic Cells

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RAW264.7 macrophages (ATCC) were cultured in Dulbecco’s Minimum Essential Media (DMEM, Life Technologies), supplemented with 10% (v/v) FBS, 1% (v/v) penicillin-streptomycin (PenStrep), and 1% (v/v) L-glutamine and cultured at 37°C and 5% CO₂. Bone marrow derived dendritic cells (BMDC) were isolated from wild-type C57/BL6 mice (Taconic) as described previously.[32 (link)] Briefly, mice were euthanized by CO₂ inhalation and femurs and tibiae were collected and sterilized in 70% (v/v) ethanol. Marrow cavity plugs were washed out using PBS. BMDC were resuspended in DMEM (Sigma Aldrich) and supplemented with 10% (w/v) bovine calf serum, 100 unit/mL penicillin, 100 µg/mL streptomycin, 2 mM L-glutamine, and 15 ng/mL granulocyte macrophage colony-stimulating factor (GM-CSF), plated onto 150-mm dishes and cultured at 37°C and 10% CO₂. Three days post-isolation, 80% of the non-adherent cells were removed and centrifuged at 300 g for 5 minutes at room temperature and fresh medium was added. Five days post-isolation, loosely adherent BMDC were collected and resuspended at 1x10⁶ cells/mL. Cells were used between 7 and 10 days post-isolation.

Lee K.L., Shukla S., Wu M., Ayat N.R., El Sanadi C.E., Wen A.M., Edelbrock J.F., Pokorski J.K., Commandeur U., Dubyak G.R, & Steinmetz N.F. (2015). Stealth filaments: polymer chain length and conformation affect the in vivo fate of PEGylated potato virus X. Acta biomaterialia, 19, 166-179.

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