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Isolation and characterization of endothelial cells from bovine cerebral arteries

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Summary

For our laboratory's investigation into the role of the endothelial cells in vasospasm following subarachnoid hemorrage and in inflammatory diseases, we found it necessary to dvise a modified method of cell culture, which would be appropriate for studying human endothelial cells from lobectomized brain. We report our techniques to increase cell harvest and ensure reproducibility, our method of culturing endothelial cells from bovine major cerebral arteries, and our morphologic and immunocytochemical charcterization of thee cells. To increase the harvest of endothelial cells, the blood cells were washed from the lumen of the major cerebral arteries at the slaughterhouse and a modified reversed vessel technique was employed. The monolayer of cultured endothelial cells displayed a cobblestone appearance when it reached confluency and transmission electron microscopy revealed junctional complexes and interdigitation of cytoplasm at Passages 10 and 17. The cells stained positively for Factor VIII-related antigen at Passages 3, 5, 7, 10, and 15. Also the cells metabolized acetylated low-density lipoprotein at Passage 3. To determine th purity of the cultured endothelial cells, an immunocytochemical study of the cytoskeleton was performed on Passage 5 cells using either rhodamine-phalloidin or antibodies against smooth muscle myosin, desmin, and vimentin.

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Authors and Affiliations

  1. Department of Internal Medicine, University of Virginia Health Sciences Center, 22908, Charlottesville, Virginia

    W. Michael Scheld

  2. Department of Neurological Surgery, University of Virginia Health Sciences Center, Box 212, 22908, Charlottesville, VA

    Takao Machi, Neal F. Kassell & W. Michael Scheld

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  1. Takao Machi
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  2. Neal F. Kassell
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  3. W. Michael Scheld
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Machi, T., Kassell, N.F. & Scheld, W.M. Isolation and characterization of endothelial cells from bovine cerebral arteries. In Vitro Cell Dev Biol 26, 291–300 (1990). https://doi.org/10.1007/BF02624460

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  • DOI: https://doi.org/10.1007/BF02624460

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