PASMCs of patients with idiopathic PAH (IPAH-PASMCs) was established as previously reported and used in the present study.39 (link) For the control experiment, PASMCs of normal subjects (Normal-PASMCs) were purchased from Lonza (Walkersville, MD). The cells were cultured in Medium 199 supplemented with 10% heat-inactivated fetal bovine serum, 100 U/mL penicillin plus 100 µg/mL streptomycin (Invitrogen/GIBCO, Grand Island), 50 µg/mL D-valine (Sigma-Aldrich) and 20 µg/mL endothelial cell growth supplement (BD Biosciences, Franklin Lakes) at 37°C. The cells were used in the experiments at passages 5 to 10. Three independent lines of IPAH- and Normal-PASMCs were used respectively.
Normal pasmcs
Manufactured by Lonza
Normal-PASMCs are primary human pulmonary artery smooth muscle cells (PASMCs) that are isolated from healthy donors. They are used for in vitro studies related to the pulmonary vasculature.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Endothelial cell growth supplement (ecgs), by BD (2 mentions)
D valine, by Merck Group (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Ckx41 microscope, by Olympus (1 mentions)
Tryple express, by Thermo Fisher Scientific (1 mentions)
3 protocols using normal pasmcs
1
Isolation and Culture of PAH Patient PASMCs
2
Isolation and Culture of PAH Patient PASMCs
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PASMCs of patients with idiopathic PAH (IPAH-PASMCs) was established as previously reported and used in the present study.39 (link) For the control experiment, PASMCs of normal subjects (Normal-PASMCs) were purchased from Lonza (Walkersville, MD). The cells were cultured in Medium 199 supplemented with 10% heat-inactivated fetal bovine serum, 100 U/mL penicillin plus 100 µg/mL streptomycin (Invitrogen/GIBCO, Grand Island), 50 µg/mL D-valine (Sigma-Aldrich) and 20 µg/mL endothelial cell growth supplement (BD Biosciences, Franklin Lakes) at 37°C. The cells were used in the experiments at passages 5 to 10. Three independent lines of IPAH- and Normal-PASMCs were used respectively.
3
Isolation and Culture of PASMCs from CTEPH Patients
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PASMCs were isolated as previously described from endarterectomized tissue removed during PEA from three patients with CTEPH.6 All experiments were carried out after approval by the Institutional Review Board of the National Hospital Organization Okayama Medical Center (approval number, H23‐RINKEN‐30). Written informed consent was obtained from each patient before the procedure. PASMCs from CTEPH patients (CTEPH PASMCs) and normal subjects (normal PASMCs; Lonza; catalog number, CC‐258; lot numbers, 0000578443, 0000658401, and 0000669096) were cultured on collagen type I‐coated dishes (Iwaki/AGC Techno Glass Co., Ltd.) in Dulbecco's modified Eagle medium (DMEM) (low glucose [1 g/L]; Gibco/Thermo Fisher Scientific) containing 10% (v/v) fetal bovine serum (FBS) with 1% (v/v) penicillin‐streptomycin (Gibco/Thermo Fisher Scientific). PASMCs were incubated in a humidified 5% CO2 atmosphere at 37°C. After reaching confluence, the cells were subcultured by trypsinization with TrypLE Express (Gibco/Thermo Fisher Scientific). Cell images were acquired with an Olympus CKX41 microscope (Olympus).
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