TDK conference 2010

The blood-brain barrier (BBB) is composed of endothelial cells, pericytes and astrocytes, and serves as interface between the blood flow and the central nervous system (CNS). Since BBB deregulation plays important role in the pathogenesis of several CNS diseases spanning from brain tumors to stroke, understanding the molecular mechanisms controlling BBB functions might help unveiling new therapeutic targets to brain pathologies. Cell-culture-based models have greatly contributed to our knowledge on the physiology, pathology and pharmacology of the blood–brain barrier (BBB). In this present study we were focusing on a novel blood–brain barrier model using primary rat brain endothelial cells, pericytes and astrocytes. To mimic ischemic events, which occur in the CNS in vivo, we applied oxygen-glucose deprivation (OGD) on BBB cells in triple co-culture and separately on the EC, PC and AC cell cultures as well. Endothelial cells appeared to be highly susceptible to cell death, whereas astrocytes and pericytes were more resistant. The results of the OGD treatment were investigated by enzyme-histochemical methods on light and electron microscopic levels, LDH release assay was performed in order to measure cell damage. A semi-quantitative assay highlighted increased ecto-ATPase activity following exposure to OGD in the three types of cell, both when grown separately and in triple co-culture. On endothelial cell cultures we also investigated cytotoxicity by hydrogen peroxide treatment, results were evaluated by statistical analysis. Taking into account our findings that are consistent with earlier in vivo experiment results, we conclude that this new in vitro model is useful for demonstrating the changes in the BBB during ischemic events and could serve as a tool for pharmacological experiments.