In vitro culture and characterization of gene targeted mouse endothelium

Endothelial cells play a crucial role in maintaining cardiovascular homeost asis. Although many cardiovascular disorders involve endothelial cell dysfu nction, the specific cellular and molecular mechanisms involved are not wel l known. We sought to establish a reproducible method of endothelial cell i solation from gene targeted mice to specifically examine endothelial pathop hysiological mechanisms. Primary aortic endothelial cell cultures were esta blished from wild type and intercellular adhesion molecule-1 (ICAM-1) defic ient mice. Isolation of mouse aortic endothelial cells (MAEC) by fluorescen t activated cell sorting routinely resulted in pure, homogenous, primary cu ltures. Wild type and ICAM-1 deficient endothelial cell morphology was simi lar, with both cultures showing cobblestone morphology and Dil-Ac-LDL stain ing. Monocyte adhesion to ICAM-1 deficient aortic endothelial cells was dec reased by 86% as compared with wild type MAEC. Monocyte adhesion was also d etermined using YN-1, an ICAM-1 blocking antibody. YN-1 decreased monocyte adhesion to wild type aortic endothelial cells by 25%, whereas YN-1 did not further decrease monocyte adhesion to ICAM-1 deficient MAEC. These data de monstrate that gene targeted endothelial cell cultures are an effective mea ns of identifying specific cellular and molecular mechanisms involved in en dothelial cell physiology and dysfunction.