DSS was purchased from MP Biomedicals (Santa Ana, CA, USA), AOM from Sigma-Aldrich (St. Louis, MO, USA), capecitabine tablets from Chia TaiTianqing Pharmaceutical Co. Ltd (Nanjing, China), Iscove’s Modified Dulbecco’s Medium (IMDM) from Sigma-Aldrich (St. Louis, MO, USA), fetal bovine serum (FBS) from Gibco (Grand Island, NY, USA), lipopolysaccharide (LPS) from Sigma Chemical Co. (St. Louis, MO, USA), recombinant Human TGF-β1 from Whiga (PeproTech Inc., USA), bicinchoninic acid (BCA) assay kits from Beyotime Biotechnology (Beijing, China), phycoerythrin (PE)-conjugated anti-CD11b, FITC-conjugated anti-Gr-1 and purified mouse Fc block (anti-CD16/CD32) from ThermoFisher Inc (Waltham, MA, USA)，Enzyme-Linked Immunosorbent Assays (ELISA) kits for IL-17, TGF-β, IL-6, tumor necrosis factor-α (TNF-α) and IL-1β from eBioscience (Waltham, MA, USA). NF-κB p-p65, p-STAT3, E-cadherin, snail, vimentin, and β-actin antibodies were purchased from Abcam, Inc (Cambridge, MA, USA). Goat Anti-rabbit IgG/Alexa Fluor 488 and Goat Anti-mouse IgG/Alexa Fluor 546 were purchased from Whiga (PeproTech Inc, USA). Goat Anti-rabbit IgG/HRP antibody was purchased from ZhongShanJinQiao Biological Technology Co (Beijing, China). Hoechst 33342 was purchased from ThermoFisher Inc (MA, USA). JSH23 was purchased from TargetMol((MA, USA).
Iscove s modified dulbecco s medium imdm
Manufactured by Merck Group
Sourced in United States, United Kingdom, Germany
Iscove's Modified Dulbecco's Medium (IMDM) is a cell culture medium formulated for the growth and maintenance of various cell types, including hematopoietic and myeloid cell lines. It is a modification of Dulbecco's Modified Eagle's Medium (DMEM) and is designed to provide a balanced salt solution and necessary nutrients for cellular proliferation and differentiation.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
L glutamine, by Thermo Fisher Scientific (5 mentions)
Streptomycin, by Thermo Fisher Scientific (5 mentions)
Penicillin, by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Merck Group (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Dulbecco s modified eagle medium dmem, by Merck Group (3 mentions)
Penicillin, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Roche (2 mentions)
Transferrin, by Merck Group (2 mentions)
Stem cell factor (scf), by R&D Systems (2 mentions)
Interleukin 3 (il 3), by R&D Systems (2 mentions)
Op9 cells, by American Type Culture Collection (2 mentions)
Erythropoietin, by Amgen (2 mentions)
Estradiol, by Pfizer (2 mentions)
Insulin, by Eli Lilly (2 mentions)
L glutamine, by Merck Group (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
2 mercaptoethanol, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Lonza (2 mentions)
30 protocols using iscove s modified dulbecco s medium imdm
1
Colorectal Cancer Mouse Model Protocol
2
Isolation and Culture of NANT Cells
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NANT cells were isolated as previously described [8 ]. Briefly, PBMCs were isolated from peripheral blood using Ficoll-Paque™ PLUS density gradient centrifugation (GE HealthCare, Uppsala, Sweden). Adherent cells were depleted by adherence, and T cells in the non-adherent fraction of PBMCs were removed using CD3+ MicroBeads according to the manufacturer’s instructions (Miltenyi Biotec, California, USA). The NANT cells were then incubated in Iscove’s Modified Dulbecco’s Medium (IMDM; Sigma-Aldrich) supplemented with 30% FBS (Sigma-Aldrich) and 1% BSA in a humidified incubator, at 37∘C with 5% CO2, for 3 days in the presence or absence of treatments. NANT cells were counted on a haemocytometer after Trypan blue staining.
3
Mouse and Human Mesenchymal Stem Cell Isolation
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Mouse bone marrow-derived MSCs (mMSCs) were obtained from the femoral and tibial bone marrow of 7–8 weeks old Balb/c mice as previously reported [60]. The protocols were approved by the Taipei Veterans General Hospital Institutional Animal Care and Use Committee (IACUC). All studies involving animals were in accordance with appropriate guidelines. mMSCs were maintained in Low-Glucose DMEM (LG-DMEM; Invitrogen) with 10% fetal bovine serum, and 1% PSG. Commercially available human MSCs (hMSCs) (Steminent Biotherapeutics Inc, Taipei, Taiwan) were maintained in Iscove’s modified Dulbecco’s medium (IMDM; Sigma-Aldrich), supplemented with 10% fetal bovine serum, 10 ng/ml fibroblast growth factor, 10 ng/ml epidermal growth factor (R&D systems, Inc.) and 100 U Penicillin-1000 U Streptomycin-2 mM L-glutamine (1% PSG; Sigma-Aldrich).
4
Erythroid Differentiation Protocol
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For determination of globin chain composition, supernatant and monolayer cells were collected at day 7 and 17 of culture, and differentiated toward the erythroid lineage as previously described (Fujita et al., 2016 (link); Uchida et al., 2017 (link)). Briefly, cells were cultured on irradiated OP9 cells (American Type Culture Collection (ATCC)) in Medium B with the addition of 5 U/mL erythropoietin (EPO; Amgen) and 5 ng/mL interleukin-3 (IL3; R&D Systems). Two days later, the floating cells were transferred into freshly irradiated OP9 feeder plates using an erythroid expansion media based on Iscove’s Modified Dulbecco’s Medium (IMDM; Sigma Aldrich) supplemented with 10 ng/mL stem cell factor (SCF; R&D Systems), 1 ng/mL IL3, 2 U/mL EPO, 1 μM estradiol (Pfizer), 1 μM dexamethasone (VETone, Boise), and 20% Knockout Serum Replacement (KSR, Thermo Fisher Scientific). Five days later, the medium was changed to a maturation erythroid medium containing IMDM, 2% bovine serum albumin (BSA; Roche), 0.56 mg/mL transferrin (Sigma Aldrich), 2 mM L-glutamine (Thermo Fisher Scientific), 2 U/mL EPO, 10 ng/mL insulin (Lilly), and 20% KSR, and the cells were cultured for another 8 to 10 days.
5
Murine Stromal Cell Culture and Differentiation
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All cells were cultured at 37 °C in a humidified atmosphere containing 5 % carbon dioxide. The SR4987 murine stromal cell line was established from a long-term bone marrow-derived cell culture of BDF/1 mice (CRL-2028; ATCC). This cell line shows a high degree of stemness [18 (link)], expresses vimentin, CD44, CD73, CD105, CD106, Sca1, and CD34, contains 50 % of CD45+ cells, and is capable of differentiating into osteocytes and chondrocytes [18 (link), 19 (link)]. Cells were grown in Iscove’s modified Dulbecco’s medium (IMDM; Sigma, St. Louis, MO, USA) supplemented with 5 % fetal bovine serum (Gibco by Life Technologies, Waltham, MA, USA) under standard conditions. U87MG GBM cells (HTB14; ATCC) were cultured in Dulbecco’s modified medium (high-glucose Dulbecco’s modified Eagle’s medium (DMEM); Sigma, St.Louis, MO, USA) supplemented with 10 % fetal bovine serum (Gibco) under standard conditions.
6
Murine Colon Cancer Cell Culture
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Murine MC38 and MC38-OVA colon cancer cells (provided by Mark Smyth, Peter MacCallum Cancer Centre, Melbourne, Australia) were cultured in Dulbecco’s modified Eagle’s medium (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Gibco), sodium pyruvate (Sigma), penicillin/streptomycin (Gibco), 2 mM L-glutamine (Sigma), and minimal essential medium nonessential amino acids (Sigma). B16-OVA cells (provided by Karine Breckpot, Vrije Universiteit Brussel, Brussels, Belgium) were cultured in Iscove’s modified Dulbecco’s medium (IMDM; Sigma) supplemented with 10% FBS (Gibco), 2 mM L-glutamine (Amimed), and a combination of 50 units per mL penicillin and 50 μg/mL streptomycin (Gibco). The cell lines used in this study were not found in the database of commonly misidentified cell lines that is maintained by the International Cell Line Authentication Committee and the National Center for Biotechnology Information. The cell lines have not been authenticated recently, but their growth behavior in vitro and in vivo was compatible with their identity. All cell lines regularly tested negative for Mycoplasma contamination.
7
Diverse Pseudomonas aeruginosa Strains for Research
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The following P. aeruginosa strains were used in this study: P. aeruginosa PAO1 (Holloway 1955 (link)), P. aeruginosa ΔpvdA ΔpchD PAO1 (ΔΔPAO1) (Visca et al. 2013 (link)) and P. aeruginosa IST27 (Leitão et al. 1996 (link)). PAO1 is a commonly used laboratory strain, whereas IST27 is a mucoid strain isolated from a cystic fibrosis patient. Other species that were used are Escherichia coli BW25113 (Grenier et al. 2014 (link)) and Salmonella enterica serovar Typhimurium NTB6 (Kortman et al. 2015 (link)). Bacteria were grown as indicated in lysogeny broth (LB; 10 g/L tryptone, 5 g/L yeast extract, 10 g/L NaCl) or Iscove’s Modified Dulbecco’s Medium (IMDM; Sigma-Aldrich). For planktonic growth experiments, pre-cultures were grown overnight in a shaking incubator at 37 °C, diluted 100-fold in fresh medium and growth was continued at 37 °C. For biofilm experiments, IMDM was supplemented with 0.5% glucose, and the biofilms were grown as indicated below.
8
Characterization of BVDV-1a Isolate MRI103
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The BVDV isolate (MRI103) used for the experimental exposures was isolated from the serum of a Scottish PI bovid which was free of maternal antibodies, and passaged six times on bovine turbinate (BT) cells. After three passages, the virus was titrated on BT cells and a multiplicity of infection (MOI) of 0.01 was used for the following passages as previously described (Frolich and Streich, 1998 (link)). Medium from the sixth passage, containing BVDV at a titre of 106 TCID50/mL, was clarified by centrifugation at 4000 × g for 30 min and stored in aliquots at -80°C before use. All cells, tissue culture medium (Iscove’s modified Dulbecco’s medium, IMDM; Sigma–Aldrich, Dorset, UK) and foetal bovine serum (FBS) used were tested free of pestivirus and antibodies against pestivirus. The 5′UTR and Npro coding region of the isolate were sequenced for phylogenetic typing as previously described (Bachofen et al., 2013b (link)) and MRI103 was determined to be a BVDV-1a virus.
9
Cell Culture Conditions for Cell Lines
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All cell lines were incubated at 37 °C in a humidified atmosphere containing 5% CO2. U2OS and HEK293FT cells were cultured in Dulbecco’s modified Eagle medium (DMEM; Sigma-Aldrich) supplemented with 10% (v/v) foetal bovine serum (FBS; BioSera), 100 U ml−1 penicillin (Gibco), 100 μg ml−1 streptomycin (Gibco) and 2 mM L-glutamine (Gibco), and for HEK293FT with 0.5 mg ml−1 G418 (Invitrogen). HAP1 cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM; Sigma-Aldrich) supplemented as above. RPE-1 cells were cultured in DMEM and F-12 Ham mix medium (Sigma-Aldrich) supplemented as above and buffered with 0.2% sodium bicarbonate (Life Technologies).
10
Erythroid Differentiation Protocol
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