Human pulmonary artery endothelial cells (HPAECs), obtained from Lonza Group (Switzerland), were maintained in complete endothelial cell growth medium-2 (Lonza) at 5% CO2 and 37 °C. The cells of three-to-six passages were used in the experiments. The HPAECs were stimulated by recombinant human TGFβ1 (R&D Systems, 5 ng/ml), IL-1β (R&D Systems, 0.1 ng/ml) and TNF-α (R&D Systems, 5 ng/ml) with or without paeoniflorin (10 μM) treatment 10 (link). For detecting intercellular signaling transduction, cells were harvested 30 min after TGFβ1, IL-1β and TNF-α challenge, while for other detections, cells were harvested 48 h after TGFβ1, IL-1β and TNF-α challenge.
Complete endothelial cell growth medium 2
Manufactured by Lonza
Sourced in Switzerland
Complete endothelial cell growth medium-2 is a cell culture medium designed to support the growth and maintenance of endothelial cells. It provides the necessary nutrients and growth factors required for the in vitro cultivation of endothelial cells.
Automatically generated - may contain errors
Lab products found in correlation
Hpaecs, by R&D Systems (1 mentions)
Recombinant human tgf β1, by R&D Systems (1 mentions)
Il 1β, by R&D Systems (1 mentions)
Tnf α, by R&D Systems (1 mentions)
Hpaec, by Lonza (1 mentions)
Collagen 1 coated, by Corning (1 mentions)
Dulbecco s phosphate buffered saline, by Thermo Fisher Scientific (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
2 protocols using complete endothelial cell growth medium 2
1
Paeoniflorin modulates inflammatory signaling in HPAECs
2
Isolation and Culture of Endothelial Colony-Forming Cells
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ECFC were isolated from venous peripheral blood following a protocol adapted from Martin‐Ramirez et al.21 A total volume of 24 mL was collected per individual, and PBMC were separated using Ficoll‐Paque PLUS (GE Healthcare Life Sciences, Pittsburgh, PA) density gradient following 30 minutes 300g centrifugation at room temperature. PBMC were further washed twice with Dulbecco's phosphate‐buffered saline (Gibco by Life Technologies, Burlington, ON, Canada) and subsequently plated on collagen I‐coated (Corning, Corning, NY) 25 cm2 tissue culture–treated Falcon flasks (Thermo Fisher Scientific, Waltham, MA) at a density of 5.0×106 cells/flask. Cultured cells were maintained at 37°C, 21% O2 and 5% CO2, using complete endothelial cell growth medium‐2 (Lonza, Basel, Switzerland) supplemented with 1% penicillin/streptomycin (Gibco by Life Technologies) and 10% fetal bovine serum. Media was changed every 2 or 3 days, and cells were maintained for up to 30 days in culture. PBMC cultures were observed daily from days 7 to 30 to determine the first day of cobblestone‐patterned ECFC colony formation. ECFC colonies were then passaged and further expanded under similar conditions. ECFC function was assessed using cells from the second passage.
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