title: Purification of endothelial cells from rat brain.
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.
authors: Department of Neurology, Garrison Institute on Aging, Texas Tech University Health Sciences Center, Lubbock, TX, USA.
authors: Jinhua
authors: J
authors: Luo
authors: Xiangling
authors: X
authors: Yin
authors: Alma
authors: A
authors: Sanchez
authors: Debjani
authors: D
authors: Tripathy
authors: Joseph
authors: J
authors: Martinez
authors: Paula
authors: P
authors: Grammas
mesh_terms: D000818:Animals; D001921:Brain / Q000098:blood supply; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology; D055806:Microvessels / Q000166:cytology; D061251:Primary Cell Culture; D051381:Rats