Complete endothelial cell growth medium mv

Human pulmonary microvascular endothelial cells (HPMECs) were purchased from PromoCell (six different batches originating from six different donors) (Heidelberg, Germany). After the cells were thawed according to the provider recommendations, the cells were maintained in complete endothelial cell growth medium MV (PromoCell) at 37 °C in a 5% CO₂ humidified atmosphere incubator. All experiments were performed using cells at passages 2 or 3. Endothelial cells were stimulated with 10 μg of isolated NETs in the presence or absence of 25 μg/ml hLR12 for 6 h.

Human pulmonary microvascular endothelial cells (HPMEC) were purchased from Promocell (6 different batches originating from 6 different donors) (Heidelberg, Germany). The cells were maintained in complete endothelial cell growth medium MV (Promocell) at 37°C in a 5% CO₂ humidified atmosphere incubator. All experiments were performed between passages 2 and 5.
Cells were stimulated in complete medium supplemented with 1 μg/ml Escherichia coli LPS (0111:B4; Sigma-Aldrich Saint-Quentin Fallavier, France) in the presence or absence of 250 or 500 nM SLC during various times depending on the experiments. Supernatants were collected for cytokines measurements and cells lysed for protein phosphorylation analyses.
Supernatants from stimulated cells were recovered after 24 h stimulation, and the concentrations of IL-8 and MCP-1 were measured using human Quantikine ELISA kits (R&D Systems, Abingdon, UK) according to the manufacturer's protocol. Cytokine array was performed using the Proteome Profiler kit (R&D Systems).