DEVELOPMENT OF AN IN VITRO HUMAN BLOOD-BRAIN BARRIER

THIS PROJECT WILL DEVELOP A WORKING IN VITRO BLOOD-BRAIN BARRIER MODEL WITH ENDOTHELIAL CELLS ISOLATED FROM HUMAN BRAIN MICROVASCULATURE. THIS MODEL WILL ALLOW RAPID ASSESSMENT OF THE EFFICACY OF DRUG DELIVERY SYSTEMS DESIGNEDTO BRING PHARMACEUTICALLY USEFUL COMPOUNDS TO THE BRAIN PARENCHYMA. OBSERVED SPECIES DIFFERENCES IN BRAIN CAPILLARY ENDOTHELIAL CELLS-POTENTIAL TARGETS FOR DELIVERY MOLECULES-PROVIDE THE RATIONALE FOR DEVELOPING A MODEL BASED ON HUMAN CELLS. PHASE I EFFORTS WILL FIRST AIM TO DEVELOP TECHNIQUES TO ISOLATE, CULTURE, AND CHARACTERIZE BRAIN CAPILLARY ENDOTHELIAL CELLS FROM FRESH HUMAN TISSUE. ONCE CULTURES ARE SET UP, EFFORTS WILL BE DIRECTED TOWARD DETERMINING CONDITIONS THAT INDUCE OR MAINTAIN BLOOD-BRAIN BARRIER-ASSOCIATED PROPERTIES IN THESE CELLS IN VITRO. THESE CHARACTERISTICS INCLUDE EXPRESSION OF APPROPRIATE BIOCHEMICAL MARKERS AND TRANSPORT PROTEINS. CELLS WILL BE GROWN ON POROUS MEMBRANES OR SOLID SUBSTRATES. EFFORTS TO OPTIMIZE THE BARRIER WILL INVOLVE EXAMING A VARIETY OF CULTURE CONDITIONS, INCLUDING THE PRESENCE OF ASTROCYTES OR THEIR PRODUCTS.