Caco-2, a human colon carcinoma cell line, was cultured in DMEM media (Welgene, South Korea) supplemented with 10% fetal bovine serum (Welgene) and 1X penicillin–streptomycin (Welgene). The cells were maintained in a humidified atmosphere of 5% CO2 and 95% air at 37°C. Upon reaching 90% confluency, the cells were seeded into 12-well plates at a density of 5 × 104 cells/well (SPL, South Korea). The culture medium was replaced every other day, and the cells were differentiated over a period of 10 days. After differentiation, bacterial cells were added to each well at concentrations of 107, 108, and 109 CFU/well in DMEM media containing 10% fetal bovine serum, followed by a 24-h incubation under standard conditions. The cytotoxicity was assessed using lactate dehydrogenase (LDH) assay (EZ-LDH, DoGenBio, South Korea) conducted using the cell supernatant according to the manufacturer’s instructions. Basal media-treated cells served as the negative control, while E. coli O157 ATCC 48395 was used as the positive cytotoxic control strain.
Dmem media
Manufactured by Welgene
15 citations
Sourced in Cameroon
About the product
DMEM (Dulbecco's Modified Eagle's Medium) is a widely used cell culture medium. It is formulated to provide the necessary nutrients and growth factors to support the proliferation and maintenance of various cell types in vitro. The medium contains essential amino acids, vitamins, salts, and glucose, providing a balanced environment for cell growth and development.
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Check compatibility
15 protocols using «dmem media»
1
Caco-2 Cell Cytotoxicity Assay
2
Culturing Lung Cancer Cell Lines
Human lung cancer cell lines, NCI-H1793, NCI-H2009, and HCC827 were purchased from the Korean Cell Line Bank (KCLB), and NCI-H1792 was purchased from the American Type Culture Collection (ATCC). NCI-H1793 and HCC827 were cultured in RPMI 1640 media (WelGENE) supplemented with 10% Fetal bovines serum (FBS; WelGENE) and antibiotics, which is comprised of 100 units/ml of penicillin, 100 μg/ml of streptomycin, and 0.25 μg/ml of Fungizone (Life Technologies Corp.). NCI-H1792 and NCI-H2009 were cultured in DMEM media (WelGENE) with the same supplements. All cells were cultured in an incubator with 5% CO2 at a temperature of 37°C.
3
Culturing Glioblastoma and Temporal Lobe Cells
Two cell lines were enzymatically dissociated to single cell from mechanically dissected glioblastoma and temporal lobe tissues. The cells were cultured in DMEM media (Welgene, Korea) supplemented with 10% fetal bovine serum (FBS; Gibco Invitrogen, USA), 100 U/mL of penicillin, and 100 mg/mL of streptomycin (Gibco Invitrogen) at 37 °C in an atmosphere of 5% CO2 in air. The cells were prepared from early passage less than 20 times and stocked (within 2 months) Additional file 2 : Fig. S1).
Corresponding organizations : Korea University, Seoul National University, Korea Institute of Science and Technology, Hanbat National University, Advanced Institute of Convergence Technology
4
Peptide-Mediated Drug Delivery for Cancer
N-terminal acylated Phe-Arg-Arg-Gly (FRRG) peptide was purchased from Peptron Co. (Daejeon, Republic of Korea). Dimethyl sulfoxide (DMSO), Dimethylformamide (DMF), doxorubicin hydrochloride (DOX), protease inhibitor cocktail, N,N-diisopropylethylamine (DIPEA), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) were purchased from Sigma Aldrich (St. Louis, MO, USA). Cathepsin B, Cathepsin E, Cathepsin D, Cathepsin L, caspase-3 and TUNEL assay kit were purchased from R&D systems (Minneapolis, MN, USA). Cathepsin B siRNA and mono-clonal Cathepsin B antibody were purchased from SantaCruz Biotechnology (Dallas,Texas, USA). Annexin V-Cy5 kit, RIPA buffer, streptavidin–horseradish peroxidase (streptavidin-HRP), BCA protein quantification kit, CellLight™ Early Endosomes-RFP, BacMam 2.0 and CellLight™ Lysosomes-RFP, BacMam 2.0 were purchased from Thermo Fisher Scientific Inc. (Rockford, IL, USA). Cell counting kit-8 (CCK-8) was purchased from Vitascientific (Beltsville, MD, USA). TEM grid (Carbon Film 200 Mesh copper) was purchased from Electron Microscopy Sciences (PA, USA). RPMI 1640 and DMEM media, antibiotics (streptomycin and penicillin) and fetal bovine serum (FBS) were purchased from WELGENE Inc. (Daegu, Republic of Korea). HT29 (human colon adenocarcinoma), MDA-MB231 (human breast adenocarcinoma), KPC960 (human pancreatic ductal adenocarcinoma) and H9C2 (rat BDIX heart myoblast) cell lines were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA).
Corresponding organizations : Korea Institute of Science and Technology, Korea University, Ewha Womans University
5
Targeted Delivery of MMAE via Ac-FRRG Peptide
N-terminal acylated FRRG (Ac-Phe-Arg-Arg-Gly-COOH) and FRRG (NH2-Phe-Arg-Arg-Gly-COOH) peptides were purchased from Peptron (Daejeon, Korea). Monomethyl auristatin E (MMAE) was purchased from MedChemExpress (Monmouth Junction, NJ, USA) Dimethylformamide (DMF), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), N,N-diisopropylethylamine (DIPEA) and N-hydroxysuccinimide (NHS) were purchased from Sigma Aldrich (St. Louis, MO, USA). TUNEL assay kit and cathepsin B enzyme were purchased from R&D systems (Minneapolis, MN, USA). CellLight™ Lysosomes-RFP, BacMam 2.0 was purchased from Thermo Fisher Scientific Inc. (Rockford, IL, USA). TEM grid (Carbon Film 200 Mesh copper) was purchased from Electron Microscopy Sciences (Atlanta, GA, USA). DMEM media, fetal bovine serum (FBS), penicillin, and streptomycin were purchased from WELGENE Inc. (Daegu, Korea). The 4T1 (murine breast adenocarcinoma) and H9C2 (rat BDIX heart myoblast) cells were purchased from ATCC (American Type Culture Collection; Manassas, VA, USA).
Corresponding organizations : Seoul National University, Korea Institute of Science and Technology, Korea University, Inje University Haeundae Paik Hospital, Ewha Womans University
Top 5 most cited protocols using «dmem media»
1
Cell Culture and Transfection Protocols
Cell Culture and Transfection MCF7 cells were maintained in RPMI media (WelGENE) supplemented with 9% fetal bovine serum (WelGENE). HEK293T and HCT116 cells were maintained in DMEM media (WelGENE) supplemented with 9% fetal bovine serum. In the cell-splitting experiments with MCF7 and HCT116 cell lines, confluent cells were split to seed 2-to 8 3 10 5 cells per 60 mm dish (21.5cm 2 of growth area, SPL Life Sciences) by treating the cells with 0.05 mM porcine trypsin and 0.53 mM EDTA (WelGENE) for 1 min. We confirmed the complete detachment of cells from each other by inspecting them through microscopic observation while counting cell numbers.
For plasmid transfection into HEK293T, the calcium-phosphate method was used and 24 hr posttransfection, the cells were split into different densities (2-to 8 3 10 5 cells per 60 mm dish). For the transfection experiment using synthetic oligonucleotides, a liposome-based method was used with Lipofectamine 2000 reagent (Invitrogen) according to the manufacturer's protocol. Six hours later, cells were split into low (2 3 10 5 cells per 60 mm dish) or high (8 3 10 5 cells per 60 mm dish) density, and RNA was prepared at indicated time points.
For plasmid transfection into HEK293T, the calcium-phosphate method was used and 24 hr posttransfection, the cells were split into different densities (2-to 8 3 10 5 cells per 60 mm dish). For the transfection experiment using synthetic oligonucleotides, a liposome-based method was used with Lipofectamine 2000 reagent (Invitrogen) according to the manufacturer's protocol. Six hours later, cells were split into low (2 3 10 5 cells per 60 mm dish) or high (8 3 10 5 cells per 60 mm dish) density, and RNA was prepared at indicated time points.
Corresponding organizations : Seoul National University
2
Antioxidant and Anti-apoptotic Assays
2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and ferric reducing/antioxidant power (FRAP), alpha glycosidase and tripydyltriazine (TPTZ) were purchased from Sigma-Aldrich (St. Louis, MO, USA). DMEM media was purchased from Welgene (Namcheon, Gyeongsan, Korea). Fetal bovine serum (FBS) and antibiotics were obtained from Gibco BRL (Rockville, MD, USA). Hybond-polyvinylidene difluoride (PVDF) membranes and electrochemiluminescence (ECL) solution were purchased from Millipore (Merck KGaA, Darmstadt, Germany). H 2 O 2 and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) were acquired from Sigma-Aldrich. SOD detection kit was purchased from Dojindo (Rockville, MD, USA). MDA detection kit was obtained from Oxis Health Products, Inc. (N. Cutter Circle, Portland, USA). A protein extraction solution (NE-PER ™ Nuclear and Cytoplasmic Extraction Reagents) was purchased from Thermo Scientific (Carlsbad, CA, USA). The following reagents were obtained from Cell Signaling Technology (Danvers, MA, USA): anti-Bcl-2, anti-Bax, anti-Cytochrome C, anti-pro-caspase 3, anti-cleavedcaspase 3, anti-pro-caspase 8, anti-cleaved-caspase 8, anti-pro-caspase 9, anti-cleaved-caspase 9, anti-PARP, anti-p38, anti-ERK, anti-JNK, anti-phospho-PI3K, anti-Total-PI3K, anti-phospho-AKT, anti-Total-AKT, antiphospho-NF-κB, anti-Nrf2, anti-lamin B, anti-β-actin, anti-HO-1, anti-NQO1 antibody and pharmacological inhibitors (LY294002 and Wortmannin). Secondary antibodies (HRP-conjugated anti-mouse IgG Ab and HRP-conjugated anti-rabbit Ab) were obtained from Calbiochem (San Diego, CA, USA).
Corresponding organizations : Korea Atomic Energy Research Institute, Kongju National University
3
Cytotoxicity and Apoptosis of BSA-CK Nanoparticles
All the cell lines were purchased from the American Type Culture Collection and the Korean Cell Line Bank (Seoul, South Korea). A549 (human lung cancer) and HT-29 (human colon cancer) cells were grown in RPMI 1640 culture medium (GenDEPOT Inc., Barker, TX), and HepG2 cells were grown in DMEM media (Welgene, Gyeongsangbuk-do, South Korea); both media were supplemented with 10% fetal bovine serum (FBS) and 1% penicillin G and streptomycin (Gibco-BRL, Gaithersburg, MD) at a temperature of 37 C in a humidi-ed incubator with a 5% CO 2 atmosphere. Cell viability was determined by MTT (3,4,5-dimethylthiazol-2-yl-2-5diphenyletrazolium bromide) assay. Cells were seeded in 96-well plates at a density of 1 Â 10 5 cells mL À1 . Aer 24 hours of incubation, the cells were treated with various concentrations of BSA-CK NPs and standard CK for 24 hours. Aer the incubation time was complete, 10 mL of MTT stock solution (5 mg mL À1 ) was added to each well and incubated for 4 hours. Then, the supernatants were removed and replaced with 100 mL of DMSO. The amount of formazan formed by the viable cells was measured using a multi-model plate reader (Bio-Tek Instruments, USA) at a test wavelength of 570 nm with a reference wavelength of 630 nm. In addition, cytotoxicity studies were conducted on HaCaT skin cell lines to analysis the viability of the cells. 25 Hoechst nuclear staining Hoechst 33 258 staining was performed to capture the apoptotic induction of BSA-CK NPs on A549 cells. Cells were seeded into a 6-well plate at a density of 1 Â 10 5 cells well À1 in 2 mL medium and incubated at 37 C with 5% CO 2 overnight. The cells were then treated with 15 mM of standard CK and BSA-CK NPs. Aer the treatment time was complete, the cells were washed twice with 1Â PBS, xed with 3.7% (v/v) formaldehyde for 5 min at room temperature, and washed twice with PBS. To dye the nucleus, the cells were stained with Hoechst 33 258 solution (2 mg mL À1 ) for 20 min in dark conditions at room temperature. The nuclear morphologies of the Hoechst-positive cells were observed and photographed under a uorescence microscope (Â400, Optinity, Korean Labtech, South Korea) for further analysis. 25 The scale bar (10 mm) was added using ImageJ soware.
Corresponding organizations : Government of the Republic of Korea, Kyung Hee University, Yong In University
4
Inflammasome Activation in Macrophages
Progenitor cells from the bone marrow were isolated from the femur and tibia of C57BL/6 mice (Nara Biotech, Seoul, Korea) and differentiated into macrophages for seven days in RPMI 1640 medium (Welgene Inc., Gyeongsan-si, Korea) containing 10% fetal bovine serum (FBS, VWR International, Wayne, PA, USA), antibiotics (CA005, GenDEPOT, Inc., Barker, TX, USA), and 30% of L929 cell-conditioned media containing macrophage colony-stimulating factor. The Raw 264.7 cell line (#40071, Korean Cell Line Bank, Seoul, Korea) was cultivated in DMEM media (Welgene Inc.) containing 10% FBS and antibiotics. All cells were incubated at 37 °C in a 5% CO2 atmosphere. To activate inflammasome (Figure 1 A), bone marrow-derived macrophages (BMDM) were plated into a culturing plate (1.0 × 106 cells per well in 12-well-plate, SPL Life Science Co., Seoul, Korea), and then primed with lipopolysaccharide (LPS, L4130, Sigma–Aldrich Co., MO, USA) for 3 h in RPMI 1649 containing 10% FBS and antibiotics. After LPS priming, the cells were subjected to RPMI 1649 medium containing inflammasome triggers as follows: adenosine triphosphate (ATP, Invitrogen, CA, USA) for 1 h; nigericin (NG, 40 μM; 4312, Tocris Bioscience, Bristol, UK) for 1 h; dsDNA (1 μg/mL) with jetPRIME (2 μL/mL, Polyplus-transfection Inc., Illkirch, France) for 1 h [15 (link)]; monosodium urate crystals (MSU, 400 μg/mL; U2875, Sigma–Aldrich Co.) which were prepared according to a previous study [16 (link)] for 3 h; flagellin (500 ng/mL, InvivoGen) with Lipofectamine 2000 (10 μL/mL, Invitrogen, Carlsbad, CA, USA) for 3 h. For the inflammasome inhibitor experiment (Figure 2 B), LPS-primed BMDM were activated by NLRP3 inflammasome and NG or MSU in the presence of KCl (50 mM, Biosesang, Seoul, Korea), Z-VAD-FMK (10 μL/mL, R&D Systems, Minneapolis, MN, USA), or MCC950 (200 nM, Invivogen). For transcripts analysis, BMDM or Raw 264.7 cells were seeded onto a 6-well plate (2.0 × 106 cells per well, SPL Life Science Co.), and treated with LPS (10 ng/mL) for 3 h with NG (10 μM), as indicated in Figure 3 A.
Corresponding organizations : Kangwon National University
5
Cell Line Acquisition and RT-PCR Analysis
T‐47D and MDA‐MB‐231 cells were obtained from the Korean Cell Line Bank (Seoul, Korea). All other cell lines were obtained from Y‐Biologics (Daejeon, Korea). MDA‐MB‐231 cells were grown in DMEM media (Welgene, Korea), while the others were maintained in RPMI‐1640 media. RT‐PCR amplification was performed using a monoplex RT‐PCR with 2X TOPsimple™ DyeMix‐multi HOT premix (Enzynomics, Korea).
Corresponding organizations : Chungnam National University, Korea Research Institute of Bioscience and Biotechnology, Korea Basic Science Institute
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