Long-term serial cultivation of arterial and capillary endothelium from adult bovine brain |
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Authors: | Ingeburg E Goetz Jean Warren Carmen Estrada Eugene Roberts Diana N Krause |
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Institution: | (1) Department of Neurobiochemistry, Beckman Research Institute of the City of Hope, 91010 Duarte, California;(2) Present address: Departamento de Fisiologia, Facultad de Medicina, Universidad Autonoma, Arzobispo Morcillo 1, 34 Madrid, Spain;(3) Department of Pharmacology, College of Medicine, University of California, 92717 Irvine, California |
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Abstract: | Summary Cerebrovascular endothelial cells from adult bovine brain were carried successfully in long-term, serial culture. Endothelial
cells were obtained from the middle and anterior cerebral arteries and from capillaries isolated from grey matter of the cerebral
cortex or caudate nucleus. Capillary cells were found to grow best in RPMI 1640 with 20% fetal bovine serum. They did not
require tumor-conditioned medium or matrix-coated surfaces, although fibronectin was used to enhance the initial plating efficiency
of the primary cultures. The same conditions were used to support satisfactory growth of arterial endothelial cells; however
they did not grow as rapidly as the capillary cells. Retention of endothelial-specific characteristics were shown for capillary-derived
cells carried up to Passage 28, arterial-derived cells up to Passage 11, and after frozen storage of both types of cultured
cells. Cultures of both arterial and capillary cells stained positively for Factor VIII antigen, exhibited a nonthrombogenic
surface, and produced prostacyclin in response to arachidonic acid. Arterial endothelial cells produced more prostacyclin
than capillary endothelium. The capillary cells had a unique tendency to assume a ringlike morphology after subculture and
sometimes formed capillarylike networks of cell cords in dense cultures. When cultured in a three-dimensional plasma clot,
capillary and arterial endothelial cells, but none of the other cell types studied, organized into tubelike structures reminiscent
of capillary formation in vivo. The availability of long-term cultures of cerebrovascular endothelial cells provides an opportunity
to compare properties of arterial and capillary endothelium from the same tissue and to investigate such processes as angiogenesis
and blood-brain barrier induction.
This work was supported in part by Public Health Service grant NS-18586 from the National Institute of Neurological and Communicative
Disorders and Stroke and by a grant from The Council for Tobacco Research—U.S.A., Inc. C. E. was supported by the Universidad
Autonoma of Madrid and the Ministerio de Universidades e Investigacion of Spain. |
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Keywords: | endothelial cell culture long-term culture cerebral artery cerebral capillaries prostacyclin angiogenesis |
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