Abstract
Endothelial cells make up a minor population of cells in a tissue, but play a major role in tissue homeostasis, as well as in diverse pathologies. To understand the biology of cerebral endothelium, purification and characterization of the cerebrocortical endothelial cell population is highly desirable. For this purpose, rat brains are mechanically minced and subsequently digested enzymatically with collagenase. In this protocol, the capillary fraction (microvessels) and the fraction enriched in small arterioles and arteries (resistance vessels) are separated. Each produces its own homogenous endothelial culture, namely, MV-EC and RV-EC. The endothelial origin of these cells is identified by positive immunofluorescent staining for the endothelial cell surface antigen Factor VIII. Unlike MV-EC, RV-EC cultures are capable of serial cultivation for up to 15 passages. Primary MV-ECs are able to retain their characteristic endothelial morphology for 6–8 weeks.
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Acknowledgments
Sources of support: This work was supported in part by grants from the National Institutes of Health (AG020569 and AG028367). Dr. Grammas is the recipient of the Shirley and Mildred Garrison Chair in Aging.
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Luo, J., Yin, X., Sanchez, A., Tripathy, D., Martinez, J., Grammas, P. (2014). Purification of Endothelial Cells from Rat Brain. In: Milner, R. (eds) Cerebral Angiogenesis. Methods in Molecular Biology, vol 1135. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0320-7_29
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DOI: https://doi.org/10.1007/978-1-4939-0320-7_29
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0319-1
Online ISBN: 978-1-4939-0320-7
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