UNLABELLED: Most in vitro studies of capillary permeability focus on endothelial cell (MVEC) monolayers and ignore the second cell that forms the capillary wall: the microvascular pericyte (PC). We describe a model to study the permeability characteristics of MVEC, PC, and MVEC:PC cocultures. METHODS: Semipermeable culture inserts were coated with collagen and then plated with early passage bovine pulmonary MVEC. On Day 3, bovine pulmonary PC were added at concentrations to approximate MVEC:PC ratios of 1:1, 5:1, and 10:1. Electrical resistance was measured on subsequent days and fluorescently labeled (FITC) albumin was used in a permeability assay to calculate an albumin clearance for each culture. RESULTS: The results for electrical resistance measurements and albumin assays showed a similar pattern. Resistance for endothelial cell monolayers was significantly higher and albumin permeability was significantly lower than that of controls. Addition of pericytes at a 10:1 and 5:1 ratios increased the permeability barrier compared to endothelial cells alone, although these cultures were not significantly different from one another. Cocultures at a 1:1 ratio had the best barrier, significantly better than all other cultures. CONCLUSIONS: Endothelial cell monolayers are an inadequate model of the microcirculation. As PC form a key component of the capillary wall in vivo and as addition of PC to MVEC monolayers in vitro, optimally at a 1:1 ratio, increase their barrier effect to large and small molecules, we believe it is necessary to include both cells in future in vitro studies. Copyright 2001 Academic Press.
UNLABELLED: Most in vitro studies of capillary permeability focus on endothelial cell (MVEC) monolayers and ignore the second cell that forms the capillary wall: the microvascular pericyte (PC). We describe a model to study the permeability characteristics of MVEC, PC, and MVEC:PC cocultures. METHODS: Semipermeable culture inserts were coated with collagen and then plated with early passage bovine pulmonary MVEC. On Day 3, bovine pulmonary PC were added at concentrations to approximate MVEC:PC ratios of 1:1, 5:1, and 10:1. Electrical resistance was measured on subsequent days and fluorescently labeled (FITC) albumin was used in a permeability assay to calculate an albumin clearance for each culture. RESULTS: The results for electrical resistance measurements and albumin assays showed a similar pattern. Resistance for endothelial cell monolayers was significantly higher and albumin permeability was significantly lower than that of controls. Addition of pericytes at a 10:1 and 5:1 ratios increased the permeability barrier compared to endothelial cells alone, although these cultures were not significantly different from one another. Cocultures at a 1:1 ratio had the best barrier, significantly better than all other cultures. CONCLUSIONS: Endothelial cell monolayers are an inadequate model of the microcirculation. As PC form a key component of the capillary wall in vivo and as addition of PC to MVEC monolayers in vitro, optimally at a 1:1 ratio, increase their barrier effect to large and small molecules, we believe it is necessary to include both cells in future in vitro studies. Copyright 2001 Academic Press.
Authors: Sara S Nunes; Laxminarayanan Krishnan; Carter S Gerard; Jacob R Dale; Melissa A Maddie; Richard L Benton; James B Hoying Journal: Microcirculation Date: 2010-10 Impact factor: 2.628
Authors: Martin Krueger; Wolfgang Härtig; Andreas Reichenbach; Ingo Bechmann; Dominik Michalski Journal: PLoS One Date: 2013-02-26 Impact factor: 3.240
Authors: Julie A Siegenthaler; Youngshik Choe; Katelin P Patterson; Ivy Hsieh; Dan Li; Shou-Ching Jaminet; Richard Daneman; Tsutomu Kume; Eric J Huang; Samuel J Pleasure Journal: Biol Open Date: 2013-05-20 Impact factor: 2.422