Immunomagnetic separation as a final purification step of liver endothelial cells |
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Authors: | Daniel E Gomez Jacqueline L Hartzler Robert H Corbitt Alexander M Nason Unnur P Thorgeirsson |
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Institution: | (1) Office of the Director, Division of Cancer Etiology, National Cancer Institute, National Institutes of Health, 9000 Rockville Pike, Building 37, Room 2D-02, 20892 Bethesda, Maryland |
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Abstract: | Summary We describe a fast and reproducible method that can be used as a final step in obtaining pure populations of liver endothelial
cells. This method employs endothelial cell specific lectin covalently bound to magnetic polystyrene beads (Dynabeads). Evonymus
europaeus agglutinin (EEA)-coated Dynabeads were used to purify monkey liver endothelium from Percoll gradient separated nonparenchymal
cells. EEA-coated beads were also successfully used to purify monkey aortic endothelial cells. The endothelial cells grew
to confluence as a cobblestonelike monolayer, expressed Factor VIII related antigen, and incorporated acetylated-low density
lipoprotein. The magnetic beads seemed not to modify the normal properties of the isolated endothelium, thus facilitating
their use in experimental studies. This immunomagnetic separation technique may be applicable for purification of endothelial
cells from a wide variety of tissue sources. |
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Keywords: | monkey liver endothelium immunoisolation culture |
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