title: Spheroid-based human endothelial cell microvessel formation in vivo.
abstract: The study of angiogenic endothelial cells (ECs) has in recent years greatly stimulated multiple fields of vascular biology research. A number of cellular models and numerous complex developmental, manipulatory and tumor animal models have been developed to study angiogenesis in vitro and in vivo. To connect the versatility of cellular assays with the complexity of readouts of in vivo experimentation, we have developed an endothelial transplantation assay. This assay is based on grafting ex vivo generated EC spheroids (2 d) in a suitable matrix in immunocompromised mice, to give rise to a 3D network of capillaries (20 d). This vasculature connects to the mouse vasculature, is perfused and matures by recruiting mouse mural cells. Here we describe the detailed protocol for this assay, including generation of spheroids, injection into mice, excision and processing of resulting plugs, and quantification by immunohistochemical analysis of the resulting vasculature.
authors: Joint Research Division Vascular Biology of the Medical Faculty Mannheim (CBTM), University of Heidelberg, Heidelberg, Germany.
authors: Anna M
authors: AM
authors: Laib
authors: Arne
authors: A
authors: Bartol
authors: Abdullah
authors: A
authors: Alajati
authors: Thomas
authors: T
authors: Korff
authors: Holger
authors: H
authors: Weber
authors: Hellmut G
authors: HG
authors: Augustin
mesh_terms: D000818:Animals; D002196:Capillaries / Q000254:growth & development*; D018929:Cell Culture Techniques / Q000379:methods*; D042783:Endothelial Cells / Q000637:transplantation*; D006801:Humans; D016867:Immunocompromised Host; D007150:Immunohistochemistry; D051379:Mice; D018919:Neovascularization, Physiologic / Q000502:physiology*; D018874:Spheroids, Cellular / Q000637:transplantation*