Novel flow cytometric analysis of the blood-brain barrier.

摘要: The blood-brain barrier (BBB) is primarily comprised of brain microvascular endothelial cells (BMVEC) and astrocytes serves as a physical chemical that separates the periphery from brain. We describe flow cytometric method using our in vitro model human BBB to characterize BMVEC surface junctional proteins critical for maintenance function, cell viability, leukocyte adhesion. For this methodology, are cocultured with transwell tissue culture insert establish barrier, after which time treated specific agents, collected analyses. use standard optimized recover coculture maintains protein expression viability. A novel adhesion assay enables quantitative analysis peripheral blood mononuclear (PBMC) interactions can be used assess many types BBB. Furthermore, concomitant large number molecules tight junction on both adherent PBMC under homeostatic pathologic conditions. Flow cytometry an extremely powerful tool, technique also applied variables not performed study, including cycle progression, calcium flux.