Ibrahim Muhammad, Richardson Michael K
Institute of Biology Leiden, Leiden University, The Netherlands; Institute of Biotechnology and Genetic Engineering, The University of Agriculture Peshawar, Pakistan.
Institute of Biology Leiden, Leiden University, The Netherlands.
Reprod Toxicol. 2017 Jun;70:102-115. doi: 10.1016/j.reprotox.2017.02.008. Epub 2017 Feb 10.
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.
组织和器官培养的一个主要限制是体外缺乏功能性血管网络。斑马鱼具有许多有用的特性，这使其成为此类研究的一个有前景的模型。不幸的是，从该物种培养内皮细胞的方法尚未得到充分表征。在这里，我们尝试了两种方法（胚状体培养和来自转基因斑马鱼kdrl:GFP胚胎的器官外植体）来构建体外血管网络。在kdrl:GFP品系中，内皮细胞表达绿色荧光蛋白，这使得能够在活细胞培养中追踪血管发育。我们发现，当在微流控系统中培养时，胚状体显示出连接的内皮细胞分支明显更长、更宽，比在静态培养中更明显。同样，在三维水凝胶基质中观察到kdrl:GFP细胞从组织外植体中发芽。这项研究是朝着体外构建斑马鱼血管网络迈出的一步。
