In Vitro Blood-Brain Barrier Model

All reagents were purchased from Millipore-Sigma (Burlington, MA, USA), unless otherwise stated. We obtained hBMECs from Neuromics (Minneapolis, MN, USA; #HEC02) [111 (link)]; these cells have been proved to be the most suitable human cell line to reproduce the BBB in vitro [112 (link)]. Adult human lung microvascular endothelial cells (HMVEC-Ls) were obtained from Lonza (Basel, Switzerland; catalog number, CC-2527) and human umbilical vein endothelial cells (HUVECs) from ThermoFisher Scientific (Waltham, MA, USA; Catalog number, #C0035C). Cells were cultured in a standard humidified atmosphere (37 °C) containing 5% CO₂, as we previously described [111 (link),113 (link)]. In some experiments, cells were transfected with pcDNA3.1-TIM-1 plasmids (GenScript, Piscataway, NJ, USA).