Immortalized mouse brain endothelial cells are ultrastructurally similar to endothelial cells and respond to astrocyte-conditioned medium |
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Authors: | John A Wijsman Richard R Shivers |
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Institution: | (1) Department of Zoology, University of Western Ontario, N6A 5B7 London, Ontario, Canada |
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Abstract: | Summary Studies of brain microvessel endothelial cell physiology and blood-brain barrier properties are often hampered by the requirement
of repeatedly producing and characterizing primary endothelial cell cultures. The use of viral oncogenes to produce several
immortalized brain microvessel cell lines has been reported. The resulting cell lines express many properties of the blood-brain
barrier phenotype but do not completely mimic primary endothelial cells in culture. As immortalized brain microvessel endothelial
cell lines have not yet been produced from mice, we transformed mouse brain endothelial cells with the adenovirus E1A gene
using a retroviral vector (DOL). Eight of 11 clones produced exhibited an endothelial-like cobblestone morphology and were
characterized as endothelial with a panel of antibodies, lectins, and ultrastructural criteria. These cells are endothelial
in origin and share ultrastructural features with primary cultures of endothelial cells. Examination of freeze fracture and
transmission electron micrographs show adherens junctions exist between the transformed cells, and culture in astrocyte-conditioned
medium induces the formation of gap junctions. This is one indication that responses to astrocyte-derived factors are retained
by the transformed cell lines. |
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Keywords: | blood-brain barrier freeze fracture ultrastructure astrocyte-conditioned medium |
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