In vitro Three-Dimensional Sprouting Assay of Angiogenesis using Mouse Embryonic Stem Cells for Vascular Disease Modeling and Drug Testing

Journal of Visualized Experiments

Abstract

Recent advances in induced pluripotent stem cells (iPSC) and gene editing technologies enable the development of novel human cell-based disease models for phenotypic drug discovery (PDD) programs. Although these novel devices could predict the safety and efficacy of investigational drugs in humans more accurately, their development to the clinic still strongly rely on mammalian data, notably the use of mouse disease models. In parallel to human organoid or organ-on-chip disease models, the development of relevant in vitro mouse models is therefore an unmet need for evaluating direct drug efficacy and safety comparisons between species and in vivo and in vitro conditions. Here, a vascular sprouting assay that utilizes mouse embryonic stem cells differentiated into embryoid bodies (EBs) is described. Vascularized EBs cultured onto 3D-collagen gel develop new blood vessels that expand, a process called sprouting angiogenesis. This model recapitulates key features of in vivo sprouting angiogenesis-formation of blood vessels from a pre-existing vascular network-including endothelial tip cell selection, endothelial cell migration and proliferation, cell guidance, tube formation, and mural cell recruitment. It is amenable to screening for drugs and genes modulating angiogenesis and shows similarities with recently described three-dimensional (3D) vascular assays based on human iPSC technologies.

Authors 4

1. Georgios Galaris Aachen Einthoven Laboratory for Experimental Vascular Medicine Department of Internal Medicine

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Einthoven Laboratory for Experimental Vascular Medicine, Department of Internal Medicine (Nephrology), Leiden University Medical Center

2. Jérémy H. Thalgott Aachen Einthoven Laboratory for Experimental Vascular Medicine Department of Internal Medicine

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Einthoven Laboratory for Experimental Vascular Medicine, Department of Internal Medicine (Nephrology), Leiden University Medical Center

3. Eliott Teston Aachen Einthoven Laboratory for Experimental Vascular Medicine Department of Internal Medicine

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Einthoven Laboratory for Experimental Vascular Medicine, Department of Internal Medicine (Nephrology), Leiden University Medical Center

4. Franck Lebrin Aachen Einthoven Laboratory for Experimental Vascular Medicine Department of Internal Medicine

   Centre National de la Recherche Scientifique · Leiden University · Université Paris Sciences et Lettres · Inserm · Leiden University Medical Center · Physique pour la médecine Paris · ESPCI Paris

   Affiliation as printed

   Einthoven Laboratory for Experimental Vascular Medicine, Department of Internal Medicine (Nephrology), Leiden University Medical Center; Institute Physics for Medicine Paris, INSERM U1273, ESPCI Paris, CNRS FRE 2031; MEMOLIFE Laboratory of Excellence and PSL Research University; F.Lebrin@lumc.nl

   Einthoven Laboratory for Experimental Vascular Medicine, Department of Internal Medicine (Nephrology), Leiden University Medical Center

   Institute Physics for Medicine Paris, INSERM U1273, ESPCI Paris, CNRS FRE 2031

   MEMOLIFE Laboratory of Excellence and PSL Research University