An in vitro three-dimensional coculture model of cerebral microvascular angiogenesis and differentiation |
| |
Authors: | Laura R. Ment William B. Stewart Dominick Scaramuzzino Joseph A. Madri |
| |
Affiliation: | (1) Department of Pediatrics, Yale University School of Medicine, 333 Cedar Street, 06510 New Haven, Connecticut;(2) Department of Neurology, Yale University School of Medicine, 333 Cedar Street, 06510 New Haven, Connecticut;(3) Department of Pathology, Yale University School of Medicine, 333 Cedar Street, 06510 New Haven, Connecticut;(4) Department of Surgery, Yale University School of Medicine, 333 Cedar Street, 06510 New Haven, Connecticut |
| |
Abstract: | Summary The microvasculature of the developing brain is plastic and responds differently to the many insults associated with preterm birth. We developed three-dimensional in vitro culture models for the study of the responses of the developing cerebral micro-vasculature. Beagle brain microvascular endothelial cells (BBMEC) were isolated by differential centrifugation from newborn beagle pups on postnatal Day 1 and placed in three-dimensional culture dispersed in a collagen gel. Alternatively, BBMEC were placed in a three-dimensional coculture with neonatal rat forebrain astrocytes. Cultures were analyzed for extracellular matrix components at 1 and 6 d, and total RNA was extracted for Northern analyses. Urokinase plasminogen activator activity was assayed in both mono- and cocultures of the two cell types. Studies of three-dimensional BBMEC/astrocyte cocultures demonstrated progressive tube formation with only low levels of endothelial proliferation. By 6 d in three-dimensional coculture, the BBMEC formed capillarylike tubes with a wrapping of glial processes, and basement membrane protein synthesis was noted. Urokinase plasminogen zymography suggested intercellular signaling by the two cell types. These data suggest that the three-dimensional beagle brain germinal matrix microvascular endothelial cell/neonatal rat astrocyte coculture provides a good model for the investigation of microvascular responses in the developing brain. |
| |
Keywords: | angiogenesis astrocytes coculture developing brain endothelial cells |
本文献已被 SpringerLink 等数据库收录! |
|