Dulbecco s modified eagle medium dmem

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Dulbecco's Modified Eagle Medium (DMEM) is a cell culture medium commonly used in laboratory research. It is a nutrient-rich solution that supports the growth and maintenance of various cell types, including adherent and suspension cells. DMEM provides essential amino acids, vitamins, and other components necessary for cell proliferation and survival.

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DMEM medium (4294 citations) Dulbecco’s modified (295 citations) Dulbeccos’s modified Eagle’s medium (DMEM; (3 citations) Dulbeccos’s modified Eagle medium (DMEM) (3 citations)

1 959 protocols using dulbecco s modified eagle medium dmem

Caffeic Acid Derivatives Induce Apoptosis

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We obtained Dulbecco's Modified Eagle Medium (DMEM) and 0.25% Trypsin from Thermo Scientific HyClone (Logan, USA). Fetal bovine serum (FBS) was purchased from Gibco (New York, USA). Dimethyl sulfoxide (DMSO) was purchased from Sangon Biotechnology Co., Ltd. (Shanghai, China). Cell counting kit-8 (CCK-8) and Apoptosis Detection Kit were obtained from the Dojindo (Kumamoto, Japan). Cell Cycle Detection Kit was purchased from KeyGEN Biotech (Nanjing, China). We purchased anti-rabbit secondary antibodies and the primary antibodies against tubulin, cytochrome C (cytoplasm), B-cell lymphoma (Bcl-2), cleaved caspase 3, P53, Bid, Bax, cleaved caspase 9, cyclin-dependent kinase 2 (CDK2), cell division control protein 2 (CDC2), cleaved caspase 8, cleaved PARP, and cyclin B1 from Abcam (Cambridge, UK). We purchased primary antibodies against phosphorylated retinoblastoma protein (P-Rb), cyclin A2, cyclin E, and E2F1 from HUABIO Biotechnology (Hangzhou, China).
Caffeic acid, p-coumaric acid, CAPE, ferulic acid, isoferulic acid, kaempferol, 3,4-dimethoxycinnamic acid, pinocembrin, naringenin, quercetin, apigenin, chrysin, and galangin were obtained from Sigma-Aldrich (Missouri, USA). Pinobanksin, 3-O-acetyl pinobanksin, and benzyl caffeate were obtained from Haishu Apexocean Biochemicals (Ningbo, China), and benzyl p-coumarate was purchased from Kunming BioBioPha Co., Ltd. (Kunming, China).

Jiang X.S., Xie H.Q., Li C.G., You M.M., Zheng Y.F., Li G.Q., Chen X., Zhang C.P, & Hu F.L. (2020). Chinese Propolis Inhibits the Proliferation of Human Gastric Cancer Cells by Inducing Apoptosis and Cell Cycle Arrest. Evidence-based Complementary and Alternative Medicine : eCAM, 2020, 2743058.

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Investigating Signaling Pathways in Cell Cultures

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4‐phenylpyridine was purchased from Sigma Aldrich (Sigma Aldrich, St. Louis, USA). Dulbecco's Modified Eagle Medium (DMEM), penicillin–streptomycin and fetal bovine serum (FBS) were from Thermo Scientific HyClone (Thermo Scientific HyClone, Logan, USA). Antibodies to β‐actin (1:1000, sc‐517,582) and c‐Src (B‐12) (1:1000, sc‐8056) were purchased from Santa Cruz Biotech (Santa Cruz Biotechnology, CA, USA). Antibodies specific for COX‐2 (1:1000, #12282), c‐Jun (1:1000, #9165), p‐c‐Jun (1:1000, #3270), α/β‐tubulin (1:1000, #2148), p38 MAPK (1:1000, #8690), phospho‐p38 MAPK (Thr180/182) (1:1000, #4511), SAPK/JNK (1:1000, #9252), phospho‐SAPK/JNK (Thr183/Tyr185) (1:1000, #4668), p44/42 MAPK (Erk1/2) (1:1000, #4695), phospho‐p44/42 MAPK (Erk1/2) (Thr202/Thr204) (1:1000, #4370), p‐Akt (S473) (1:1000, #9271), Akt (1:1000, #9272), MKK3/6 (1:1000, #9238), p‐MKK3/6 (1:1000, #9231), MKK4/7 (1:1000, #9152), p‐MKK4/7 (1:1000, #9156), MEK1/2 (1:1000, #9122), p‐MEK1/2 (1:1000, #9121), p‐Src (Y416) (1:1000, #6943), p‐EGFR (Y845) (1:1000, #2231), p‐EGFR (Y1068) (1:1000, #3777), p‐EGFR (Y1045) (1:1000, #2237) and EGFR (1:1000, #4267) were purchased from Cell Signalling Biotechnology (Cell Signalling Biotechnology,). Anti‐lamin B1 (1:10000, ab 16,048) was purchased from Abcam (Abcam).

Kim J.G., Kang H.Y., Kim M.J., Lim S., Lee C.J., Kim K, & Jung S.K. (2022). 4‐phenylpyridine suppresses UVB‐induced skin inflammation by targeting c‐Src in vitro and in vivo. Journal of Cellular and Molecular Medicine, 26(14), 3891-3901.

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CaCo-2 and HEK293T cell cultivation

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CaCo-2 cells (kindly provided by Konstantin Sparrer, University Hospital Ulm, Germany) were cultivated in DMEM (Dulbecco’s Modified Eagle Medium DMEM; 11500516, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS-12A; Capricorn Scientific, Ebsdorfergrund, Germany), 2 mM Gluta-MAX™ (35050061, Thermo Fisher Scientific), 25 mM HEPES (15630080, Thermo Fisher Scientific), 1× MEM Non-Essential Amino Acids Solution (11140050, Thermo Fisher Scientific), and 50 µg/mL gentamycin (1405-41-0, Serva Electrophoresis, Heidelberg, Germany) and passaged every 2–3 days depending on confluence.
HEK293T T7/N cells (previously described in [19 (link)]) were cultivated in DMEM (Dulbecco’s Modified Eagle Medium DMEM; 11500516, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated FBS, 2 mM Gluta-MAX™, 25 mM HEPES, 5 µg/mL blasticidin (asnt-bl-1, InvivoGen, San Diego, CA, USA), and 2 µg/mL puromycin (SC-1080713, Santa Cruz Biotechnology, Dallas, TX, USA).
All cells were incubated at 37 °C, 5% CO₂, and 80% relative humidity.

Cordsmeier A., Jungnickl D., Herrmann A., Korn K, & Ensser A. (2023). Analysis of SARS-CoV-2 Spike Protein Variants with Recombinant Reporter Viruses Created from a Bacmid System. International Journal of Molecular Sciences, 24(9), 8156.

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Cell Culture Protocols for Pancreatic Cancer

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Panc-1 and H1650 cells were purchased from ATCC (Manassas, VA, USA). bxPC-3 and MIA PaCa-2 were gifted by Dr. Diane Simeone lab (University of Michigan, Ann Arbor, MI, USA). All cells were maintained at 37 °C in 5% CO₂. Cells were grown to 70%–80% confluence, before subculturing using 0.05% Trypsin-EDTA (Thermo Fisher Scientific, Waltham, MA, USA). Between subculturing, media was replenished every 48–72 h. Panc-1 cells were grown in Dulbecco’s Modified Eagle Medium (DMEM) (Thermo Fisher Scientific), supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, St. Louis, MO, USA) and 1% Antibiotic-antimycotic (Thermo Fisher Scientific). MIA PaCa-2 cells were grown in in Dulbecco’s Modified Eagle Medium (DMEM) (Thermo Fisher Scientific), supplemented with 10% FBS (Sigma-Aldrich), 2.5% horse serum and 1% Antibiotic-antimycotic (Thermo Fisher Scientific). bxPC-3 and H1650 cells were grown in RPMI-1640 (Thermo Fisher Scientific), supplemented with 10% FBS (Sigma-Aldrich) and 1% Antibiotic-antimycotic (Thermo Fisher Scientific). Cells were routinely tested for and reported negative for mycoplasma contamination.

Rivera-Báez L., Lohse I., Lin E., Raghavan S., Owen S., Harouaka R., Herman K., Mehta G., Lawrence T.S., Morgan M.A., Cuneo K.C, & Nagrath S. (2020). Expansion of Circulating Tumor Cells from Patients with Locally Advanced Pancreatic Cancer Enable Patient Derived Xenografts and Functional Studies for Personalized Medicine. Cancers, 12(4), 1011.

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Culturing Mutu DCs, Mutu Cas9 DCs, and Cell Lines

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Mutu DCs (Fuertes Marraco et al. 2012) and Mutu Cas9 DCs (Wilson et al. 2018) were cultured in Iscove's Modified Dulbecco's Medium (IMDM; Gibco; Scoresby, Australia) supplemented with 10 % (v/v) fetal bovine serum (FBS), 60 μg/mL penicillin, 100 μg/mL streptomycin and 100 μM β-mercaptoethanol (Fuertes Marraco et al. 2012; Wilson et al. 2018) . To passage, Mutu DCs were lifted from the flask using ethylenediaminetetraacetic acid-balanced salt solution (EDTA-BSS; 150 mM sodium chloride, 4 mM potassium chloride, 24 μM disodium hydrogen orthophosphate, 12 μM sodium dihydrogen orthophosphate, 15 mM HEPES, and 5 mM EDTA (The Peter Doherty Institute for Infection and Immunity media preparation unit [MPU]; Melbourne, Australia) supplemented with 2 % (v/v) FBS). RAW264.7 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco) supplied with 10 % (v/v) FBS and 1 % (v/v) antibiotic-antimycotic. RAW264.7 cells were lifted from the flasks using a rubber policeman. HSC-3 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco) supplied with 10 % (v/v) FBS and 1 % (v/v) antibiotic-antimycotic (Momose et al. 1989) . HSC-3 cells were lifted using trypsin (1.25 %). All cells were maintained in humidified incubators at 37 °C and 5 % CO 2 .

Xu B., Anderson B.M., Mountford S.J., Thompson P.E., Mintern J.D, & Edgington-Mitchell L.E. (2024). Cathepsin X deficiency alters the processing and localisation of cathepsin L and impairs cleavage of a nuclear cathepsin L substrate. Biological chemistry, 405(5).

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SARS-CoV-2 Variant Propagation in Vero Cells

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Vero cells (CCL-81, ATCC) were maintained in Dulbecco's Modified Eagle Medium (DMEM) (Gibco, Gaithersburg, MD, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% penicillin/streptomycin (PS) (Gibco). CBNU-nCoV01 (S clade), CBNU-nCoV11 (a B.1.1.7 variant), CBNU-nCoV21 (a B.1.351 variant), and CBNU-nCoV40 (a B.1.1.529 variant) were propagated in Vero cells in Dulbecco's modified Eagle medium (DMEM; Gibco, Grand Island, NY) supplemented with 1% penicillin-streptomycin (Gibco) and TPCK (tosylsulfonyl phenylalanyl chloromethyl ketone)-treated trypsin (0.5 μg/mL; Worthington Biochemical, Lakewood, NJ) in a 37 °C incubator supplemented with 5% CO₂ for 72 h.

Kim E.H., Lee B.W., Ryu B., Cho H.M., Kim S.M., Jang S.G., Casel M.A., Rollon R., Yoo J.S., Poo H., Oh W.K, & Choi Y.K. (2022). Inhibition of a broad range of SARS-CoV-2 variants by antiviral phytochemicals in hACE2 mice. Antiviral Research, 204, 105371.

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Cell Culture of Human and Mouse Monocytes

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THP-1 cell lines (Human Monocyte Leukemia Cells) were purchased from the American Type Culture Collection (ATCC) and cultured in Roswell Park Memorial Institute (RPMI) Medium 1640 (Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone) and 1% (v/v) penicillin/streptomycin (P/S, Gibco) at 37 °C in 5% CO₂ environment. RAW 264.7 cell lines (Mouse Monocyte-macrophage Leukemia Cells) were purchased from the American Type Culture Collection (ATCC), and maintained in Dulbecco’s modified Eagle medium (DMEM) (Gibco) supplemented with 10% fetal bovine serum (FBS, Hyclone) and 1% (v/v) penicillin/streptomycin (P/S, Gibco) at 37 °C in 5% CO₂ environment. L929 cell lines (Mouse Fibroblasts Cells) were purchased from the American Type Culture Collection (ATCC), and maintained in Dulbecco’s modified Eagle medium (DMEM) (Gibco) supplemented with 10% Heat-inactivated horse serum (Gibco) and 1% (v/v) penicillin/streptomycin (P/S, Gibco) at 37 °C in 5% CO₂ environment. Human neutrophils were isolated from peripheral blood mononuclear cells (PBMCs) of healthy donors and cultured in serum-free RPMI 1640 at 37 °C in 5% CO₂ environment.

Tan Q., He L., Meng X., Wang W., Pan H., Yin W., Zhu T., Huang X, & Shan H. (2021). Macrophage biomimetic nanocarriers for anti-inflammation and targeted antiviral treatment in COVID-19. Journal of Nanobiotechnology, 19, 173.

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Eggshell-Derived Biomaterials for Osteoblast Culture

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Raw white eggshells were provided by American Dehydrated Foods, Atlanta, GA. Ethanol (99.5% purity, absolute), nitric acid (65% HNO₃), ammonium hydroxide (28% NH₄OH), semiconductor grade phosphoric acid (85% H₃PO₄), poly(lactic acid) (PLA, Mw: 76000), anhydrous chloroform (≥99% purity), and dimethyl sulfoxide (≥99% DMSO) were purchased from Sigma-Aldrich Chemical Company St. Louis, MO. Human osteoblast cells (ATCC CRL 11372) were purchased from American Type Culture Collection (ATCC); Dulbecco's modified eagle medium (DMEM) was purchased from Fisher scientific. Penicillin and streptomycin were purchased from Lonza. Fetal bovine serum (FBS Lot no. 8SB013) was purchased from VWR. Hematoxylin and eosin reagents were purchased from Sigma-Aldrich.

Apalangya V.A., Rangari V.K., Tiimob B.J., Jeelani S, & Samuel T. (2019). Eggshell Based Nano-Engineered Hydroxyapatite and Poly(lactic) Acid Electrospun Fibers as Potential Tissue Scaffold. International Journal of Biomaterials, 2019, 6762575.

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Formulation and Characterization of Lipid Nanoparticles

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1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-mPEG2k), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and cholesterol hemisuccinate were purchased from Avanti Polar Lipids. Linear poly(ethyleneimine) (2.5kD) was purchased from Polysciences. β-Cyclodextrin, pyrene and curcumin were purchased from Sigma. DNAase-I was purchased from New England Biolabs. EGFP-NLS was obtained as a gift from Prof. Rob Parton (Addgene plasmid# 67652). Slide-a-Lyzer dialysis cassettes (MWCO 20K, 3 mL) and cell culture reagents like Dulbecco’s modified Eagle medium (DMEM), Opti-MEM^® reduced serum medium, 1× phosphate buffered saline solution (PBS) and Gibco^TM TrypLE express enzyme (1×) were purchased from Fisher Scientific. Fetal bovine serum (FBS), Penicillin-Streptomycin 10/10 (100X) and L-glutamine –200 mM (100×) were purchased from Atlanta Biologicals. MB49 cells were obtained as a gift from Prof. Timothy Ratliff (Purdue University, Department of Comparative Pathobiology) and have been authenticated by STR profile. Ultrapure water (18.2 MΩ) was used for preparation of 20 mM HEPES buffer (pH 7.4). All solvents were of analytical grade, purchased from commercial sources. Buffers and solvents were filtered through 0.22 µm CA syringe filters (Macherey-Nagel Inc.) before use.

Samaddar S., Mazur J., Boehm D, & Thompson D.H. (2019). Development And In Vitro Characterization Of Bladder Tumor Cell Targeted Lipid-Coated Polyplex For Dual Delivery Of Plasmids And Small Molecules. International Journal of Nanomedicine, 14, 9547-9561.

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Investigating Molecular Pathways in Cell Migration

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Sodium dichromate dihydrate (Na₂Cr₂O₇) was from Sigma (St Louis, MO, USA). Dulbecco’s Modified Eagle Medium (DMEM) and F12/K were purchased from Fisher Sci (Waltham, MA). Transfection reagent PolyJet was purchased from SignaGen Laboratories (Rockville, MD). Primary antibodies against NEDD9, Slug, non-p-β-catenin, and β-catenin, and secondary antibodies against mouse and rabbit were purchased from Cell Signaling Inc (Beverly, MA, USA). GAPDH antibody was purchased from Gentex Corporation (Irvine, CA, USA). SuperScript first-strand synthesis kit was purchased from Invitrogen (Waltham, MA, USA). miRScript PCR kit was purchased from Qiagen (Hilden, Germany). PowerUP SYBR green master mix was purchased from Applied Biosystems (Waltham, MA). A Dual-Luciferase Assay kit was purchased from Promega (WI, USA). ECF substrate for Western blot was purchased from GE Healthcare (Pittsburgh, PA, USA).

Zhang Z., Li J., Yan B., Tu H., Huang C, & Costa M. (2022). Loss of MEG3 and upregulation of miR-145 play an important role in the invasion and migration of Cr(VI)-transformed cells. Heliyon, 8(8), e10086.

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