Isolated HPASMC were cultured in Smooth Muscle Cell Basal Medium (CC-31–81; Lonza) supplemented with 10 % FBS and growth factors from SmGM-2 SingleQuots (CC-4149; Lonza), 100 U/ml penicillin and 0.1 mg/ml streptomycin; (15140–122; Gibco). Moreover, HPASMC were isolated from pulmonary arteries obtained from patients with PAH or those with chronic obstructive pulmonary disease (COPD), undergoing lung transplantation. Mouse PASMC were isolated from the pulmonary arteries.
Smooth muscle cell basal medium
Manufactured by Lonza
Smooth Muscle Cell Basal Medium is a cell culture medium designed to support the growth and maintenance of human smooth muscle cells. It provides the necessary nutrients and growth factors required for the optimal proliferation and differentiation of these cell types.
Automatically generated - may contain errors
Lab products found in correlation
Smgm 2 singlequots, by Lonza (2 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Lab tek 2 4 well chamber slides, by Thermo Fisher Scientific (1 mentions)
Endothelial cell basal medium mv, by PromoCell (1 mentions)
Penicillin, by Lonza (1 mentions)
Streptomycin, by Lonza (1 mentions)
Hcaec, by PromoCell (1 mentions)
Endothelial cell basal medium 2, by Lonza (1 mentions)
Egm 2 mv singlequots, by Lonza (1 mentions)
Hcasmc, by Lonza (1 mentions)
Vascular endothelial growth factor (vegf), by R&D Systems (1 mentions)
Ebm 2, by Lonza (1 mentions)
Hcaec, by Lonza (1 mentions)
3 protocols using smooth muscle cell basal medium
1
HPASMC Isolation and Culture
2
Culturing Coronary Artery Endothelial and Smooth Muscle Cells
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Human coronary artery endothelial cells (HCAEC, PromoCell, Heidelberg, Germany) were cultured in Endothelial cell basal medium MV (PromoCell) supplemented with the Supplement Pack Endothelial Cell GM MV (PromoCell) comprising fetal calf serum, endothelial cell growth supplement, recombinant human epidermal growth factor, heparin, and hydrocortisone; as well as 100 U/ml penicillin and 100 μg/mL streptomycin (Lonza, Basel, Switzerland) under standard cell culture conditions (37 °C, 5% CO2). For microscopy, 6 × 104 cells were seeded into each well of Lab-Tek II 4-well Chamber slides (Nunc A/S, Roskilde, Denmark), cultured until they reached 80–95% confluency, and fixed with 4% PFA for 15 min and kept in PBS until the time of staining.
Human coronary artery smooth muscle cells (CaSMC, Clonetics, Lonza) were cultured under standard cell culture conditions (37 °C, 5% CO2) in Smooth Muscle cell Basal Medium (Clonetics Lonza) supplemented with SmGm-2 SingleQuots supplement pack containing epidermal growth factor, fibroblast growth factor, insulin, gentamicin sulfate, and fetal bovine serum; as well as 100 U/ml penicillin and 100 μg/ml streptomycin (Lonza). For microscopy, 2.5 × 104 cells were seeded into each well of Lab-Tek II 4-well Chamber slides (Nunc), cultured until they reached 80–95% confluency, and fixed with 4% PFA for 15 min and kept in PBS until the time of staining.
Human coronary artery smooth muscle cells (CaSMC, Clonetics, Lonza) were cultured under standard cell culture conditions (37 °C, 5% CO2) in Smooth Muscle cell Basal Medium (Clonetics Lonza) supplemented with SmGm-2 SingleQuots supplement pack containing epidermal growth factor, fibroblast growth factor, insulin, gentamicin sulfate, and fetal bovine serum; as well as 100 U/ml penicillin and 100 μg/ml streptomycin (Lonza). For microscopy, 2.5 × 104 cells were seeded into each well of Lab-Tek II 4-well Chamber slides (Nunc), cultured until they reached 80–95% confluency, and fixed with 4% PFA for 15 min and kept in PBS until the time of staining.
3
Stimulation of Coronary Cells
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Human coronary artery endothelial cells (HCAECs) and human coronary artery smooth muscle cells (HCASMCs) were purchased from Lonza. HCAECs and HCASMCs were grown in Endothelial Cell Basal Medium‐2 (EBM‐2; Lonza) supplemented with EGM‐2 MV SingleQuots (Lonza) and in Smooth Muscle Cell Basal Medium (Lonza) supplemented with SmGM‐2 SingleQuots (Lonza), respectively, according to the manufacturer's instructions. After serum starvation for 24 hours, cells were stimulated with 50 ng/mL of vascular endothelial growth factor (VEGF) (R&D Systems) for 0.5 or 24 hours, 0.5 μmol/L insulin for 0.5 hours, 100 μmol/L H2O2 for 1 hour followed by 23‐hour incubation in normal culture medium after washing, 0 to 200 nmol/L recombinant FABP4, or 10 μg/mL anti‐FABP4 antibody in the medium supplemented with 0.5% BSA. The doses of reagents and incubation periods varied according to the experimental protocol. Each experiment was performed in at least triplicate.
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