The synthesis of the ligands and complexes was performed according to the protocol described by Sgarbossa and co-workers [18 (link)]. Complexes and ligands were dissolved in DMSO as a 10 mM stock solution and stored at room temperature. SB202190, AZD6244, and SP600125 were purchased from Selleck Chemical (Houston, TX); they were dissolved in DMSO as a 20 mM stock solution and stored at −20 °C. Vanadyl sulfate (VOSO4), 4’,6-diamidino-2-phenylindole (DAPI), Mowiol® 4-88, RNase, propidium iodide, EDTA, protease inhibitor cocktails, and monoclonal anti-β-actin antibody were purchased from Sigma-Aldrich (St. Louis, MO). VOSO4 was freshly dissolved in medium. DMEM, DMEM-F12, RPMI 1640, and fetal bovine serum (FBS) were purchased from Aurogene (Rome, IT). EGF, insulin, hydrocortisone, penicillin/streptomycin antibiotic mixture, amphotericin B, and glutamine were purchased from Sigma-Aldrich (Milan, IT).
Dulbecco modified eagle medium (dmem)
Manufactured by Aurogene
Sourced in Italy
DMEM (Dulbecco's Modified Eagle's Medium) is a widely used cell culture medium designed to support the growth and maintenance of a variety of cell types. It contains essential nutrients, amino acids, vitamins, and salts necessary for cell proliferation and survival. DMEM is a common component in many cell culture applications and is often supplemented with additional factors, such as serum or growth factors, to meet the specific requirements of the cells being cultivated.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Aurogene (2 mentions)
Au l0022, by Aurogene (2 mentions)
Au s181h, by Aurogene (2 mentions)
Au x0550, by Aurogene (2 mentions)
Anti β actin monoclonal antibody, by Merck Group (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Insulin, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Glutamine, by Merck Group (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Mowiol 4 88, by Merck Group (1 mentions)
Penicillin streptomycin antibiotic mixture, by Merck Group (1 mentions)
Amphotericin b, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Azd6244, by Selleck Chemicals (1 mentions)
Sb202190, by Selleck Chemicals (1 mentions)
Sp600125, by Selleck Chemicals (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Euroclone (1 mentions)
6 protocols using dulbecco modified eagle medium (dmem)
1
Synthesis and Characterization of Ligand-Metal Complexes
2
Isolation and Characterization of Calvarium-Derived Mesenchymal Stromal Cells
3
Exploring Cardiac H9c2 Cell Response
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Embryonic rat cardiac H9c2 (2-1) cells (ECACC, Salisbury, UK) were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Aurogene, Rome, Italy), containing 5.5 mM d-glucose and supplemented with 10% heat-inactivated fetal bovine serum (FBS; AU-S181H Aurogene, Rome, Italy), 1% L-Glutamine (L-Glu; AU-X0550 Aurogene, Rome, Italy), and 1% penicillin/streptomycin solution (P/S; AU-L0022 Aurogene, Rome, Italy), at 37 °C under an atmosphere of 5% CO2.
After reaching 80% confluence, H9c2 cells were trypsinized, seeded at a specific cell density for each assay, and then exposed to NG, high glucose (HG; 33 mM d-glucose), or NG + 27.5 mM mannitol (M; as an osmotic control) for 48 h [40 (link)]. Cells were then treated for 6 days [41 (link)] in NG or HG medium with the following substances:
After reaching 80% confluence, H9c2 cells were trypsinized, seeded at a specific cell density for each assay, and then exposed to NG, high glucose (HG; 33 mM d-glucose), or NG + 27.5 mM mannitol (M; as an osmotic control) for 48 h [40 (link)]. Cells were then treated for 6 days [41 (link)] in NG or HG medium with the following substances:
CHR 0.399 mg/mL (CHR), dissolved in NaCl;
SBECD 7.3 m/m%, dissolved in NaCl;
Binary system SBECD + 0.095 mg/mL CHR (SBECD + CHR), dissolved in NaCl;
DMSO 2.5% as a vehicle of OTX008;
OTX008 (0.75–1.25–2.50 µM);
Binary system OTX008 (2.5–1.25–0.75 µM)-SBECD (OTX008-SBECD), dissolved in NaCl;
Ternary system CHR (0.324 mg/mL)-OTX008 (2.5–1.25–0.75 µM)-SBECD (CHR-OTX008-SBECD), dissolved in NaCl.
4
SHSY-5Y Cell Culture Protocol
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SHSY-5Y cells (kindly provided by I. Szabò) were grown in Dulbecco’s Modified Eagle Medium (DMEM, Aurogene) supplemented with 10% (v/v) fetal bovine serum (FBS), 100 U/mL penicillin G, 0.1 mg/mL streptomycin and 5% glutamine.
5
Pancreatic Cancer Cell Line Maintenance
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Pancreatic cancer cell lines (HPAFII, Aspc1, HS766T, PANC1, and BxPc3) were from ATCC (Manassas, VA, USA); Pan02 cells were from the National Cancer Institute (Frederick Cancer Research and Development Center, Frederick, MD, USA). Cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Aurogene, Roma, Italy) (Pan02, HPAFII, Aspc1, HS766T, PANC1) or in RPMI medium (BxPc3) supplemented with 10% (v/v) fetal bovine serum (FBS), 100 U/mL penicillin G, 0.1 mg/mL streptomycin, and 5% glutamine. Cells were grown at 37 °C in a humified atmosphere supplemented with 5% CO2.
6
Cardiac H9c2 Cell Glucose Exposure and Treatments
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As previously described (Hermenean et al., 2023 (link)), embryonic rat cardiac H9c2 (2-1) cells (ECACC, United Kingdom) were cultured at 37°C under an atmosphere of 5% CO2, in Dulbecco’s modified Eagle’s medium (DMEM; Aurogene, Italy). This growth medium contained 5.5 mM D-glucose, 1% L-Glutamine (L-Glu; AU-X0550 Aurogene, Italy), 10% heat inactivated fetal bovine serum (FBS; AU-S181H Aurogene, Italy) and 1% penicillin/streptomycin (P/S) solution (AU-L0022 Aurogene, Italy). H9c2 cells were seeded at a specific density for each assay before being exposed to NG, high glucose (HG; 33 mM D-glucose) or NG + 27.5 mM mannitol (M; as osmotic control) for 48 h (Hermenean et al., 2023 (link)). Cells were then treated in NG or HG medium for 6 days (Hermenean et al., 2023 (link)) with the following substances:
- CHR 0.399 mg/mL dissolved in NaCl (CHR);
- SBECD 7.3 m/m% dissolved in NaCl (SBECD);
- SBECD + 0.095 mg/mL CHR dissolved in NaCl (SBECD + CHR);
- as vehicle for OTX008, dimethyl sulfoxide 2.5% (DMSO);
- OTX008 (0.75–1.25–2.50 µM);
- SBECD-OTX008 (2.5–1.25–0.75 µM) dissolved in NaCl (SBECD + OTX);
- SBECD-OTX008 (2.5–1.25–0.75 µM)-CHR dissolved in NaCl (SBECD + OTX + CHR).
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