title: In vitro development of zebrafish vascular networks.
abstract: A major limitation to culturing tissues and organs is the lack of a functional vascular network in vitro. The zebrafish possess many useful properties which makes it a promising model for such studies. Unfortunately, methods of culturing endothelial cells from this species are not well characterised. Here, we tried two methods (embryoid body culture and organ explants from transgenic zebrafish kdrl:GFP embryos) to develop in vitro vascular networks. In the kdrl:GFP line, endothelial cells expresses green fluorescent protein, which allows to track the vascular development in live cultures. We found that embryoid bodies showed significantly longer and wider branches of connected endothelial cells when grown in a microfluidic system than in static culture. Similarly, sprouting of kdrl:GFP+cells from the tissue explants was observed in a 3D hydrogel matrix. This study is a step towards the development of zebrafish vascular networks in vitro.
authors.affiliation: Institute of Biology Leiden, Leiden University, The Netherlands; Institute of Biotechnology and Genetic Engineering, The University of Agriculture Peshawar, Pakistan.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Ibrahim
authors.affiliation: Institute of Biology Leiden, Leiden University, The Netherlands. Electronic address: m.k.richardson@biology.leidenuniv.nl.
authors.forename: Michael K
authors.initials: MK
authors.lastname: Richardson
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002478:Cells, Cultured; D004625:Embryo, Nonmammalian / Q000098:blood supply*; D058732:Embryoid Bodies / Q000502:physiology*; D042783:Endothelial Cells / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006321:Heart / Q000196:embryology; D008099:Liver / Q000166:cytology / Q000196:embryology; D009206:Myocardium / Q000166:cytology; D018919:Neovascularization, Physiologic; D015027:Zebrafish
chemical_list: D049452:Green Fluorescent Proteins
keywords: Angiogenesis; Embryoid bodies; Explant culture; Microfluidics; Vasculogenesis; Zebrafish