Human pulmonary artery smooth muscle cells (hPASMCs; Cascade Biologics Inc., Portland, OR) were cultured in SmGM-2 BulletKit media (Lonza, Basel, Switzerland) containing 5% (volume/volume) heat-inactivated fetal bovine serum (FBS; Gibco, Carlsbad, CA), 0.5 ng/ml human recombinant epidermal growth factor, 2 ng/ml human recombinant fibroblast growth factor, 5 μg/ml insulin, and 50 μg/ml gentamicin. HEK293 cells were maintained in DMEM supplemented with 10% heat-inactivated FBS, penicillin (100 U/ml), and streptomycin (100 μg/ml) at 37°C in a humidified atmosphere containing 5% CO2. Cells at passages 6–8 were used for experiments. For induction of hypoxia, cells were transferred in a special hypoxia incubator (Thermo Scientific, model 3130, Rockford, IL) with 3% O2, 5% CO2, and balanced nitrogen. The O2 concentration inside the chamber was detected continuously by using an oxygen monitor (Hudson Ventronics Division, CA) to ensure that the O2 concentration was 3%.
Model 3130
Manufactured by Thermo Fisher Scientific
The Model 3130 is a capillary electrophoresis instrument designed for DNA analysis. It features a 16-capillary array and utilizes laser-induced fluorescence detection technology. The instrument is capable of performing various electrophoretic separations, including DNA sequencing, fragment analysis, and genotyping.
Automatically generated - may contain errors
2 protocols using model 3130
1
Culturing and Hypoxic Stimulation of hPASMCs
2
ARPE-19 Cell Culture in Normoxia and Hypoxia
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The human RPE cell line ARPE‐19 (#CRL‐2302, American Type Culture Collection) was cultured in DMEM containing glutamine and supplemented with 50 U penicillin/50 mg streptomycin and 10% foetal bovine serum (FBS) (#F8687, Sigma‐Aldrich). ARPE‐19 cells in the normoxia (normal) group were cultured under 95% O2/5% CO2, and ARPE‐19 cells in the hypoxia group were cultured under 95% N2/5% CO2 in an incubator (Model 3130; Thermo Fisher Scientific).
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