title: Vascularization of human glioma spheroids implanted into rat cortex is conferred by two distinct mechanisms.
abstract: Aim of this study was to develop and characterize an applicable in vivo model to investigate angiogenesis of human gliomas. An established glioblastoma spheroid model was used to investigate the neovascularization of a standardized avascular solid tumor mass. Spheroids of two human glioma cell lines were labeled with an in vivo fluorescent dye. Single spheroids were implanted into the cortex of athymic rats. After 1, 3, 7, 14, and 21 days, brain sections containing the spheroid were immunostained for endothelial cells or vascular endothelial growth factor (VEGF). The dye-stained glioma spheroid and the endothelial cells were visualized by confocal microscopy. Two distinct mechanisms of tumor vascularization could be observed. (1) "Classical" angiogenesis with new vessels sprouting from existing host vessels into the spheroid was seen. (2) Individual endothelial cells were found to migrate towards and into the center of the spheroid where they coalesced to form new vessels. This process occurred as early as 24 hr after spheroid implantation. Spheroid vascularization was accompanied by an increase of VEGF expression, which peaked 7 days after implantation and returned to normal patterns by 14-21 days. Besides the "classical" angiogenesis by angiogenic blood vessels, the recruitment of individual endothelial cells seems to be an additional mechanism in early glioma vascularization. Our model proves to be a reliable, reproducible system to study in vivo angiogenesis of human gliomas.
authors.affiliation: Department of Neurosurgery, University of Wuerzburg, Germany. goldbrunner@mail.uni-wuerzburg.de
authors.forename: R H
authors.initials: RH
authors.lastname: Goldbrunner
authors.forename: J J
authors.initials: JJ
authors.lastname: Bernstein
authors.forename: K H
authors.initials: KH
authors.lastname: Plate
authors.forename: G H
authors.initials: GH
authors.lastname: Vince
authors.forename: K
authors.initials: K
authors.lastname: Roosen
authors.forename: J C
authors.initials: JC
authors.lastname: Tonn
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000098:blood supply* / Q000737:chemistry / Q000473:pathology; D002465:Cell Movement; D016228:Endothelial Growth Factors / Q000032:analysis; D004730:Endothelium, Vascular / Q000166:cytology; D005910:Glioma / Q000098:blood supply*; D006801:Humans; D008222:Lymphokines / Q000032:analysis; D008297:Male; D018613:Microscopy, Confocal; D009368:Neoplasm Transplantation; D009389:Neovascularization, Pathologic; D010841:Pia Mater / Q000098:blood supply; D020033:Protein Isoforms / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D018874:Spheroids, Cellular / Q000737:chemistry / Q000473:pathology / Q000502:physiology*; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
chemical_list: D016228:Endothelial Growth Factors; D008222:Lymphokines; D020033:Protein Isoforms; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors