Astrocytes induce neural microvascular endothelial cells to form capillary‐like structures in vitro

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Abstract
Astrocytes maintain a unique association with the central nervous system microvasculature and are thought to play a role in neural microvessel formation and differentiation. We investigated the influence of astroglial cells on neural microvascular endothelial differentiation in vitro. Using an astroglial‐endothelial coculture system, rat brain astrocytes and C6 cells of astroglial lineage are shown to induce bovine retinal microvascular endothelial (BRE) cells to form capillarylike structures. Light microscopic evidence for endothelial reorganization began within 48 hours and was complete 72‐96 hours following the addition of BRE cells to 1‐day‐old astroglial cultures. The extent of BRE reorganization was quantitated by computer‐assisted analysis and shown to be dependent upon the density of both the BRE and C6 cells within the cocultures. Coculture conditions in which BRE cells were separated from C6 cells by porous membranes failed to generate this endothelial cell change. Likewise, C6‐conditioned media and C6‐endothelial coculture conditioned media did not induce BRE cell reorganization. Extracellular laminin within the C6‐endothelial cocultures, identified by indirect immunofluorescence, was concentrated at the endothelial‐astroglial interface of capillary‐like structures consistent with incipient basement membrane formation. Astroglial cells accumulated adjacent to capillary‐like structures suggesting the presence of bidirectional influences between the reorganized endothelial cells and astroglia. This is the first demonstration of astroglial induction of angiogenesis in vitro and these findings support a functional role for perivascular astrocytes in the vascularization of neural tissue such as retina and brain.

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