Primary culture of capillary endothelium from rat brain |
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| Authors: | Phillip D. Bowman A. Lorris Betz Diane aR Jerry S. Wolinsky Jack B. Penney Richard R. Shivers Gary W. Goldstein |
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| Affiliation: | (1) University of Michigan, Ann Arbor, Michigan;(2) Johns Hopkins University, Baltimore, Maryland;(3) The University of Western, Ontario, London, Canada;(4) Department of Pediatrics, University Hospital, F2705B, 48109 Ann Arbor, Michigan |
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| Abstract: | Summary To provide an in vitro system for studies of brain capillary function we developed a method for culture of brain capillary endothelial cells. Capillaries were isolated from rat brain and enzymatically treated to remove the basement membrane and contaminating pericytes. Subsequent Percoll gradient centrifugation resulted in a homogeneous population of capillary endothelial cells that attached to a collagen substrate and incorporated [3H]thymidine. Evidence for the endothelial nature of these cells was provided by the presence of Factor VIII antigen and angiotensin converting enzyme activity and by the failure of platelets to adhere to the cell surface. In addition, the cells were joined together by tight junctions. Thus, primary cultures of these cells retained both endothelial and blood-brain barrier features. This study was supported by the National Foundation-March of Dimes and by Grants HL-25492 and ES-02380 from the National Institutes of Health. J. S. W. is the recipient of a Research Career Development award (NS-00443) and J. B. P. is the recipient of a Teacher-Investigator award (1P01-NS15655). |
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| Keywords: | rat brain endothelial cell culture primary culture Factor VIII angiotensin converting enzyme tight junctions |
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