Isolation of endothelial cells from murine tissue

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 dige...

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Bibliographic Details
Published inJournal of immunological methods Vol. 244; no. 1; pp. 205 - 215
Main Authors Marelli-Berg, Federica M, Peek, Emma, Lidington, Elaine A, Stauss, Hans J, Lechler, Robert I
Format Journal Article
LanguageEnglish
Published Amsterdam Elsevier B.V 20.10.2000
Elsevier
Subjects
Animals
Biological and medical sciences
CD31 antigen
Cell Culture Techniques - methods
Coronary Vessels - cytology
Diverse techniques
Endothelial cells
Endothelium, Vascular - cytology
Female
Fundamental and applied biological sciences. Psychology
Immunomagnetic Separation - methods
Isolation
Lung - blood supply
Mice
Mice, Inbred CBA
Molecular and cellular biology
Mouse
Primary cultures
Isolation
Primary cultures
Mouse
Endothelial cells
Vertebrata
Endothelial cell
Mammalia
Rodentia
Primary culture
Method
Experimental protocol
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Summary: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.
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ISSN:0022-1759
1872-7905
DOI:10.1016/S0022-1759(00)00258-1