A Matrigel assay was set up in a flat-bottomed 96-well plate in triplicates as previously described [23 (link)]. In brief, MS1 (a mouse pancreatic endothelial cell line) were trypsinized, counted and resuspended in EBM-2 basal medium (Lonza, USA). Matrigel basement membrane (Millipore, MA, USA) was plated into each well (100 μl/well) and incubated for 1 hour at 37°C until adequate polymerization. Next, 2×104 VECs (MS1) were co-cultured in basal media alone, with supplemented growth factors [5% FBS, VEGF, FGF, EGF, IGF] (EGM-2MV endothelial cell growth medium; Lonza, USA) or with 2×104 mast cells (stimulated with 1 ng/ml IL-33 for 24 hrs) (1:1 ratio; 100 μl/well) on the matrigel surface, and was incubated at 37°C.Tube formation was observed for 12 hours, and micrographs of co-cultures were captured using an inverted brightfield microscope (Leica).
Egm 2mv endothelial cell growth medium
Manufactured by Lonza
Sourced in United States
EGM-2MV endothelial cell growth medium is a complete, serum-free medium designed for the growth and maintenance of human endothelial cells. It provides the essential nutrients and growth factors required for the optimal growth of endothelial cells in culture.
Automatically generated - may contain errors
Lab products found in correlation
Ebm 2 basal medium, by Lonza (2 mentions)
Inverted bright field microscope, by Leica (1 mentions)
Matrigel basement membrane, by Merck Group (1 mentions)
Dmem f12, by Merck Group (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Gelatin, by Merck Group (1 mentions)
Fibronectin, by BD (1 mentions)
Spectramax plus 384 microplate reader, by Molecular Devices (1 mentions)
3 protocols using egm 2mv endothelial cell growth medium
1
Matrigel-Based Angiogenesis Assay with Endothelial and Mast Cells
2
Multilineage Differentiation Potential of Endothelial Cells
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Twelve-well plates were coated with a solution containing 50 μg/mL fibronectin (BD Bioscience) and 0.1% gelatin (Sigma-Aldrich). Cells were seeded with the density of 2.0 × 104 cells per well in the complete cell culture medium (DMEM/F12 supplemented with 10% FBS, Sigma-Aldrich; and 100 IU/mL penicillin, 100 µg/mL streptomycin, Gibco, ThermoFisher Scientific) for 24 h. The following day, the culture medium was changed with EGM-2MV Endothelial Cell Growth Medium (Lonza, Basel, Switzerland). EGM-2MV was replaced every two days. Cells were examined for endothelial differentiation on days 7, 14 and 21 days of culture, following staining for endothelial markers.
The expression of selected osteogenic, chondrogenic, adipogenic, cardiomyogenic and angiogenic-specific markers was evaluated by measuring the levels of corresponding mRNAs as well as by histochemical or immunocytochemical staining.
The expression of selected osteogenic, chondrogenic, adipogenic, cardiomyogenic and angiogenic-specific markers was evaluated by measuring the levels of corresponding mRNAs as well as by histochemical or immunocytochemical staining.
3
Endothelial Cell Proliferation Assay
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