Abstract Title
Differential Expression of Extracellular Matrix Genes Under Various Co-Culture Models of the Blood-Brain Barrier
Abstract
In vitro cell and tissue cultures are a powerful tool for targeted analysis of isolated cell populations. These studies, however, are hampered by phenotypic drift and an inevitable loss of physiologic relevance that results from the removal of cells from their natural multicellular signaling environment. This issue can be partially mitigated through the use of co-culturing techniques, which utilize specialized devices to culture two or more physiologically-relevant cell types. In this project, we utilized an in vitro blood-brain barrier (BBB) model to characterize expression of extracellular matrix (ECM) genes between various co-culturing of brain derived pericytes, astrocytes and endothelial cells. Modeled BBB function was determined by transwell solute-permeability assays, and ECM mRNA expression was measured using reverse-transcription qPCR. Through this process we were not only able to demonstrate that co-culturing itself enhances BBB function, but we also utilized this method to determine how different BBB co-culture models exhibit marked differences in the genes involved in regulation of the BBB-ECM. Future studies with these BBB models will allow us to determine the relative contribution of various cell-types to the ECM microenvironment, and how these environments may be altered during inflammatory conditions and disease states.
Differential Expression of Extracellular Matrix Genes Under Various Co-Culture Models of the Blood-Brain Barrier
In vitro cell and tissue cultures are a powerful tool for targeted analysis of isolated cell populations. These studies, however, are hampered by phenotypic drift and an inevitable loss of physiologic relevance that results from the removal of cells from their natural multicellular signaling environment. This issue can be partially mitigated through the use of co-culturing techniques, which utilize specialized devices to culture two or more physiologically-relevant cell types. In this project, we utilized an in vitro blood-brain barrier (BBB) model to characterize expression of extracellular matrix (ECM) genes between various co-culturing of brain derived pericytes, astrocytes and endothelial cells. Modeled BBB function was determined by transwell solute-permeability assays, and ECM mRNA expression was measured using reverse-transcription qPCR. Through this process we were not only able to demonstrate that co-culturing itself enhances BBB function, but we also utilized this method to determine how different BBB co-culture models exhibit marked differences in the genes involved in regulation of the BBB-ECM. Future studies with these BBB models will allow us to determine the relative contribution of various cell-types to the ECM microenvironment, and how these environments may be altered during inflammatory conditions and disease states.