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Dulbecco modified eagle medium (dmem)

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DMEM (Dulbecco's Modified Eagle's Medium) is a well-established cell culture medium formulation commonly used to support the growth and maintenance of various cell lines in vitro. It provides a balanced salt solution, essential amino acids, vitamins, and other nutrients required for cellular proliferation and survival.

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Lab products found in correlation

Fetal bovine serum (fbs), by Atlanta Biologicals (17 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions) Horse serum, by Atlanta Biologicals (5 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions) Hydrocortisone, by Merck Group (3 mentions) Cholera toxin, by Merck Group (3 mentions) Streptomycin, by Atlanta Biologicals (3 mentions) Dulbecco modified eagle medium (dmem), by Corning (3 mentions) Penicillin, by Thermo Fisher Scientific (3 mentions) Mda mb 231, by American Type Culture Collection (2 mentions) Human epidermal growth factor (hegf), by Merck Group (2 mentions) Bovine insulin, by Merck Group (2 mentions) Penicillin, by Merck Group (2 mentions) Penicillin g, by Atlanta Biologicals (2 mentions) Penicillin streptomycin, by Atlanta Biologicals (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Corning (2 mentions) Mcf 10a, by American Type Culture Collection (2 mentions) C2c12 myoblast, by American Type Culture Collection (2 mentions)

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DMEM medium (2 citations)

26 protocols using dulbecco modified eagle medium (dmem)

1

Organotypic Culture of Cervical Epithelial Cells

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Organotypic culture was performed as previously described47 (link). However, in these experiments we omitted epidermal growth factor (EGF) and reduced the concentration of fetal bovine serum to 5%. Collagen rafts were made in 6-well cell culture plates (Corning). Six collagen rafts were formed by mixing 2.5 ml of rat tail collagen I (Millipore), 8 ml of DMEM with 10% FBS (Atlanta Biologicals), 2 ml of DMEM with casein (5 mg/ml), 1 × 106 of 3T3-J2 cells or mitomycin C-treated human stromal cells, and 0.5 ml of 0.1 M NaOH. Two ml of this mixture was aliquoted to each well and incubated at 37 °C for 2 days to allow contraction of rafts before 1 × 106 cervical epithelial cells were allowed to attach to the top of each raft (for 2 hours). All HPV16-immortalized cervical cells were tested at approximately 80 to 95 population doublings or about 25 to 30 passages. Rafts were submerged in raft medium48 (1:3 mixture of F12/DMEM medium with 5% FBS plus Supplements) for 48 hours before being raised onto stainless steel grids in a 100 mm cell culture dish to form an air-liquid interface for epithelial cells. The medium was changed every 2 days and cultures were fixed after 10 days.
Deng H., Hillpot E., Mondal S., Khurana K.K, & Woodworth C.D. (2018). HPV16-Immortalized Cells from Human Transformation Zone and Endocervix are More Dysplastic than Ectocervical Cells in Organotypic Culture. Scientific Reports, 8, 15402.
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2

Evaluation of Tumor Cell Lines

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Karpas299 (K299) cells expressing receptor CD30 were used at the National Cancer Institute/National Institutes of Health, in collaboration with Dr. Mark Raffeld. CD30-negative U937 and MDA-MB-231 cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA). K299 and MDA-MB231 cells, stably expressing GFP and luciferase, were used for tumor formation in mouse model. Suspension cells K299 and U937 were cultured with RPMI1640 medium (Fisher Scientific, Pittsburgh, PA) with 10% FBS (Atlanta Biologicals, Lawrenceville, GA). Adhesion cells MDA-MB-231 were cultured in DMEM (Atlanta Biologicals, Lawrenceville, GA) with 10% FBS. The chemotherapeutic drug DOX was purchased from Sigma (St. Louis, MO).
Zhao N., Zeng Z, & Zu Y. (2017). Self-Assembled Aptamer-Nanomedicine for Targeted Chemotherapy and Gene Therapy. Small (Weinheim an der Bergstrasse, Germany), 14(4), 10.1002/smll.201702103.
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3

Endocannabinoid Receptor Modulation in Cell Proliferation

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The following were purchased from Cayman (Ann Arbor, MI): 2-AG; the endocannabinoid receptor inhibitors, SR141716A (Rimonabant, CB1 inverse agonist), AM251 (CB1 antagonist) and SR144528 (CB2 inverse agonist); the endocannabinoid receptor activators, CP47497 (CB1 agonist), AM1241 (CB2 agonist), Win- 55–212-2 (CB1 and CB2 agonist) and CP55940 (CB1 and CB2 agonist); the selective and nonselective inhibitors for cyclooxygenase-1 and -2 (COX-1, COX-2), SC-560 (selective for COX-1), CAY10404 (selective for COX- 2), Ibuprofen (nonselective for COX-1 and COX-2); and the selective inhibitors for hydrolases of 2-AG, JZL 184 (selective for enzyme monoacylglycerol lipase, MAGL. Anti-bromodeoxyurine (BrdU) antibody was purchased from Roche Applied Science (Indianapolis, IN). The CellTiter 96 Non-Radioactive Cell proliferation Assay Kit™ was purchased from Promega (Madison, WI). Charcoal/dextran-treated fetal bovine serum (CFBS) was purchased from Hyclone (Logan, UT). DMEM, RPMI-1640, antibiotics (penicillin and streptomycin), and fetal bovine serum (FBS) were purchased from Atlanta Biologicals (Lawrenceville, GA).
Zhang J., Medina-Cleghorn D., Bernal-Mizrachi L., Bracci P.M., Hubbard A., Conde L., Riby J., Nomura D.K, & Skibola C.F. (2016). The potential relevance of the endocannabinoid, 2-arachidonoylglycerol, in diffuse large B-cell lymphoma. Oncoscience, 3(1), 31-41.
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4

Murine Brain Cell Culture Protocol

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The complete culture medium for all cell lines was DMEM supplemented with 10% fetal bovine serum (ATLANTA Biologicals, Flowery Branch, GA, USA) and 1% Penicillin/Streptomycin solution (HyClone Laboratories Inc., South Logan, UT, USA). Murine brain cell lines bEnd.3 (endothelial cells), N2a (neuroblastoma cells), and C8-D1A (astrocytes) were purchased from ATCC. Murine brain cell line BV-2 (microglia) was obtained from Dr. G. Jean Harry (National Institute of Environmental Health Sciences, Research Triangle Park, NC). All cell lines were maintained according to provided protocols and previous studies [5 ]. Briefly, the cells were cultured in 75 cm2 flasks (Corning, NY, USA) separately, and medium was refreshed every other day until the cells reached ≥ 90% confluency.
Liu L., Koo Y., Akwitti C., Russell T., Gay E., Laskowitz D.T, & Yun Y. (2019). Three-dimensional (3D) brain microphysiological system for organophosphates and neurochemical agent toxicity screening. PLoS ONE, 14(11), e0224657.
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5

Solid-Phase Synthesis of Lipid Nanoparticles

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Fmoc-protected amino acids, resins, and coupling reagents for solid-phase synthesis were purchased from EMD Millipore. Tris(3-hydroxypropyltriazolylmethyl)amine (THPTA) was purchased from Sigma-Aldrich. 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-mPEG1000 (mPEG1000-DSPE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-PEG2000-azide (N3-PEG2000-DSPE), and cholesterol (Chol) were purchased from Avanti Polar Lipids. Cyanine 5.5-NHS ester was purchased from Lumiprobe. Cellulose acetate (CA) syringe filters were purchased from Macherey-Nagel Inc. Sephadex® G-50 and LH-20 were purchased from GE Healthcare. LIVE/DEAD® fixable green dead cell stain was purchased from ThermoFisher. AlexaFluor® 680-Wheat Germ Agglutinin conjugate (WGA) and LysoTracker® Blue DND-22 were purchased from Life Technologies. Cell culture reagents such as DMEM, FBS, trypsin, and PBS were purchased from Atlanta Biologicals. Ultrapure water (18 MΩ) was used for preparation of all buffers and in all experiments. All solvents were of analytical grade, purchased from commercial sources and used without further purification, except DMF which was dried over CaH2 under N2, filtered, and distilled under reduced pressure.
Lee Y., Kischuk E., Crist S., Ratliff T.L, & Thompson D.H. (2017). Targeting and Internalization of Liposomes by Bladder Tumor Cells Using a Fibronectin Attachment Protein-derived Peptide-Lipopolymer Conjugate. Bioconjugate chemistry, 28(5), 1481-1490.
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6

Culturing TNBC and Normal Mammary Cells

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Triple-negative breast cancer (MDA-MB-231 and CRL-2321) and normal mammary epithelial (MCF10A) cell lines were obtained from the American Type Culture Collection. MCF10A-H cells, derived via H-ras transformation of MCF10A cells, were a kind gift from Dr Barbara Stefanska, Purdue University. MDA-MB-231 cells were cultured in Dublecco's modified essential medium (DMEM; Life Technologies, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals, Norcross, GA, USA), 100 IU/ml penicillin and 100 µg/ml streptomycin (Life Technologies). MCF10A and MCF10A-H cells were cultured in a 1:1 ratio of DMEM:Ham's F12 supplemented with 5% horse serum (HS; Atlanta Biologicals), 20 ng/ml human epidermal growth factor (Sigma-Aldrich, St. Louis, MO, USA), 0.5 mg/ml hydrocortisone (Sigma-Aldrich), 100 ng/ml cholera toxin (Sigma-Aldrich), 10 µg/ml bovine insulin (Sigma-Aldrich), 100 IU/ml penicillin and 100 µg/ml streptomycin. CRL2321 cells were cultured in Roswell Park Memorial Institute 1640 medium (RPMI-1640, Mediatech Inc., Herndon, VA, USA) supplemented with 10% FBS, 100 IU/ml penicillin and 100 µg/ml streptomycin.
Raman V., Lorenzo J.L., Stashenko E.E., Levy M., Levy M.M, & Camarillo I.G. (2017). Lippia origanoides extract induces cell cycle arrest and apoptosis and suppresses NF-κB signaling in triple-negative breast cancer cells. International Journal of Oncology, 51(6), 1801-1808.
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7

Isolation and Culture of Colon Fibroblasts

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Fibroblasts were isolated from primary tissues using previously described methodologies [3 (link)]. Briefly, after mincing, heated collagenase digestion on a shaking water bath was used and the filtrate was cultured on tissue culture plates in DMEM 10% FBS (Atlanta Biologicals) with 1% Pen/Strep (Gibco). Human colon fibroblasts cell lines CRL1541, CRL1459, and CRL7213 were obtained from the American Type Culture Collection (ATCC). Primary colon fibroblasts were isolated from human colon tissue was obtained with informed consent from patients undergoing surgery without preoperative treatment for sporadic colon cancer or colectomy for chronic ulcerative colitis. Primary fibroblasts were cultured from non-dysplastic sections of colon (normal), adenocarcinoma or colitic sections as previously described [3 (link)].
Signs S.A., Fisher R.C., Tran U., Chakrabarti S., Sarvestani S.K., Xiang S., Liska D., Roche V., Lai W., Gittleman H.R., Wessely O, & Huang E.H. (2018). Stromal miR-20a controls paracrine CXCL8 secretion in colitis and colon cancer. Oncotarget, 9(16), 13048-13059.
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8

HNSCC Cell Lines and PARP Inhibitors

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Human HNSCC cell lines UMSCC1, UMSCC14, and UMSCC17 were originally from Thomas Carey at the University of Michigan and maintained in the lab [53 (link)]. Mouse embryonic fibroblasts were isolated from C57BL6 mice. Cells were cultured in DMEM supplemented with 10% fetal bovine serum (Atlanta Biologicals, Atlanta, GA) and maintained at 37°C under a humidified 5% CO2 atmosphere. Poly (ADP-ribose) polymerase (PARP) inhibitors 6(5H)-phenanthridinone (PHEN) and N-(6-Oxo-5,6-dihydrophenanthridin-2-yl)-(N, N-dimethylamino) acetamide hydrochloride (PJ34), and APR-246 were purchased from Sigma-Aldrich (St. Louis, MO). zVAD-fmk was purchased from Biomol International (Plymouth Meeting, PA).
Yin Z.X., Hang W., Liu G., Wang Y.S., Shen X.F., Sun Q.H., Li D.D., Jian Y.P., Zhang Y.H., Quan C.S., Zeng Q., Li Y.L., Zhao R.X., Ding Q, & Xu Z.X. (2017). PARP-1 inhibitors sensitize HNSCC cells to APR-246 by inactivation of thioredoxin reductase 1 (TrxR1) and promotion of ROS accumulation. Oncotarget, 9(2), 1885-1897.
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9

Culturing Glioblastoma Cells Under Normoxia and Hypoxia

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U87 and U87 EGFRvIII mutant cells (U87vIII) were provided by Dr. Nathan Price (Institute for Systems Biology, Seattle, WA). Cells were cultured in Dulbecco’s modified eagle medium (DMEM; Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS; Atlanta Biologicals, Flowery Branch, GA) and 1% penicillin/streptomycin (Lonza) at 37°C in a 5% CO2 environment and passaged upon confluence. At the start of each experiment, cells were homogeneously mixed with the pre-polymerized hydrogel solution at a density of 1 × 105 cells/25 μl hydrogel then photopolymerized as described above. Cell-seeded hydrogels were incubated in cell culture medium at 37°C, 5% CO2 in low adhesion well-plates containing standard culture media with ~20% O2 (normoxia) or 1% O2 (hypoxia) in a dedicated, computer controlled hypoxia chamber (C-Chamber Hypoxia Chamber, Biospherix™, Parish, NY) within the cell culture incubator. Media were changed at days 3 and 5 for cultures extending to 7 days. Media exchange for samples in the hypoxia chamber was performed using culture media pre-conditioned in the hypoxia chamber for at least 12 hours.
Chen J.W., Lumibao J., Blazek A., Gaskins H.R, & Harley B. (2018). Hypoxia activates enhanced invasive potential and endogenous hyaluronic acid production by glioblastoma cells. Biomaterials science, 6(4), 854-862.
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10

Cell Culture Conditions for Aptamer Selection

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The cell lines, including B lymphoma Maver-1, mantle cell lymphoma Jeko-1, Burkitt's lymphoma CA46, acute erythroid leukemia HEL, diffuse histiocytic lymphoma SU-DHL-1 and breast cancer MDA-MB-468 were purchased from ATCC (Manassas, VA). All cell suspensions were cultured in RPMI 1640 medium (Thermo Fisher Scientific, Rockford, IL) containing 10% Fetal Bovine Serum (FBS, Atlanta Biologicals, Lawrenceville, GA), and 100 IU/mL penicillin-streptomycin. All adhesion cell lines were cultured with DMEM (Atlanta Biologicals, Lawrenceville, GA) containing 10% FBS and 100 IU/mL penicillin-streptomycin. The washing buffer used during aptamer enrichment contained 4.5 g/L glucose and 5 mmol/L MgCl2 in Dulbecco's Phosphate-Buffered Saline (DPBS, Sigma Aldrich, St. Louis, MO). Binding buffer was prepared by adding Bovine Serum Albumin (1 mg/mL; BSA, Thermo Fisher Scientific) with 0.1 mg/mL t-RNA (Sigma Aldrich) to the washing buffer.
Li H., Yang S., Yu G., Shen L., Fan J., Xu L., Zhang H., Zhao N., Zeng Z., Hu T., Wen J, & Zu Y. (2017). Aptamer Internalization via Endocytosis Inducing S-Phase Arrest and Priming Maver-1 Lymphoma Cells for Cytarabine Chemotherapy. Theranostics, 7(5), 1204-1213.
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