Isolation of endothelial cells from murine tissue Export

@article{citeulike:2190074, abstract = {The isolation and long-term culture of murine endothelial cells (ECs) has often proven a difficult task. In this paper we describe a quick, efficient protocol for the isolation of microvascular endothelial cells from murine tissues. Murine lung or heart are mechanically minced and enzymatically digested with collagenase and trypsin. The single cell suspension obtained is then incubated with an anti-CD31 antibody, anti-CD105 antibody and with biotinylated isolectin B-4. Pure EC populations are finally obtained by magnetic bead separation using rat anti-mouse Ig- and streptavidin-conjugated microbeads. EC cultures are subsequently expanded and characterised. The surface molecule expression by the primary cultures of murine EC obtained from lung and heart tissue is analysed and compared to that of a murine endothelioma and of primary cultures of murine renal tubular epithelial cells. The phenotype and morphology of these cultures remain stable over 10-15 passages in culture, and no overgrowth of contaminating cells of non-endothelial origin is observed at any stage.}, author = {Marelli-Berg, Federica M. and Peek, Emma and Lidington, Elaine A. and Stauss, Hans J. and Lechler, Robert I.}, citeulike-article-id = {2190074}, citeulike-linkout-0 = {http://dx.doi.org/10.1016/S0022-1759(00)00258-1}, citeulike-linkout-1 = {http://www.sciencedirect.com/science/article/B6T2Y-41D4N4K-R/2/703d36dc71f5175af09bf62724ff54c3}, day = {20}, doi = {10.1016/S0022-1759(00)00258-1}, journal = {Journal of Immunological Methods}, keywords = {culture, endothelial\_cells, isolation}, month = {October}, number = {1-2}, pages = {205--215}, posted-at = {2008-01-02 23:02:34}, priority = {2}, title = {Isolation of endothelial cells from murine tissue}, url = {http://dx.doi.org/10.1016/S0022-1759(00)00258-1}, volume = {244}, year = {2000} }

TY - JOUR ID - citeulike:2190074 L3 - citeulike-article-id:2190074 N2 - The isolation and long-term culture of murine endothelial cells (ECs) has often proven a difficult task. In this paper we describe a quick, efficient protocol for the isolation of microvascular endothelial cells from murine tissues. Murine lung or heart are mechanically minced and enzymatically digested with collagenase and trypsin. The single cell suspension obtained is then incubated with an anti-CD31 antibody, anti-CD105 antibody and with biotinylated isolectin B-4. Pure EC populations are finally obtained by magnetic bead separation using rat anti-mouse Ig- and streptavidin-conjugated microbeads. EC cultures are subsequently expanded and characterised. The surface molecule expression by the primary cultures of murine EC obtained from lung and heart tissue is analysed and compared to that of a murine endothelioma and of primary cultures of murine renal tubular epithelial cells. The phenotype and morphology of these cultures remain stable over 10-15 passages in culture, and no overgrowth of contaminating cells of non-endothelial origin is observed at any stage. IS - 1-2 JF - Journal of Immunological Methods EP - 215 TI - Isolation of endothelial cells from murine tissue VL - 244 SP - 205 KW - culture KW - endothelial_cells KW - isolation AU - Marelli-Berg, Federica AU - Peek, Emma AU - Lidington, Elaine AU - Stauss, Hans AU - Lechler, Robert PY - 2000/10/20/ UR - http://dx.doi.org/10.1016/S0022-1759(00)00258-1 ER -