*5.4. Primary Culture*

The human glomerular endothelial cells (HGEC) were obtained as previously described by Amaral et al. [28] from kidneys of human pediatric patients, under proper consent and ethical approval (N◦: 035 LUP1S0/19). HGEC were cultivated in M199 media, supplemented with 20% fetal calf serum (FCS), 3.2 mM L-glutamine, 100 U/mL penicillin/streptomycin (GIBCO, Waltham, MA, USA), and 25 μg/mL endothelial cell growth supplement (ECGS, Sigma, St. Louis, MO, USA). All the experiments were performed with

HGEC between 2–7 passages and were previously characterized for positive expression of von Willebrand factor and platelet/endothelial cell adhesion molecule 1 (PECAM-1). Moreover, experiments were carried out at growth-arrested conditions using M199 medium supplemented with 10% FCS without ECGS [28].
