Single-cell characterization of neovascularization using hiPSC-derived endothelial cells in a 3D microenvironment

• Published in: Stem cell reports 2023-10, Vol.18 (10), p.1972-1986
• Main Authors: Rosowski, Simon, Remmert, Caroline, Marder, Maren, Akishiba, Misao, Bushe, Judith, Feuchtinger, Annette, Platen, Alina, Ussar, Siegfried, Theis, Fabian, Wiedenmann, Sandra, Meier, Matthias
• Format: Article
• Language: English
• Subjects: culturing technologies; inferred EC-pericyte interactions; microfluidic ligand assay; neovascularization; single-cell mRNA sequencing; stem cell-derived endothelial cells
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2023.08.008

The formation of vascular structures is fundamental for in vitro tissue engineering. Vascularization can enable the nutrient supply within larger structures and increase transplantation efficiency. We differentiated human induced pluripotent stem cells toward endothelial cells in 3D suspension culture. To investigate in vitro neovascularization and various 3D microenvironmental approaches, we designed a comprehensive single-cell transcriptomic study. Time-resolved single-cell transcriptomics of the endothelial and co-evolving mural cells gave insights into cell type development, stability, and plasticity. Transfer to a 3D hydrogel microenvironment induced neovascularization and facilitated tracing of migrating, coalescing, and tubulogenic endothelial cell states. During maturation, we monitored two pericyte subtypes evolving mural cells. Profiling cell-cell interactions between pericytes and endothelial cells revealed angiogenic signals during tubulogenesis. In silico discovered ligands were tested for their capability to attract endothelial cells. Our data, analyses, and results provide an in vitro roadmap to guide vascularization in future tissue engineering. [Display omitted] •In this study, we engineered stem cell-derived vessels in a 3D cell culture•Time-resolved single-cell transcriptomes reveal similarities to human embryogenesis•Discovery of cell communication between pericytes and ECs during tubulogenesis In this article, Meier and colleagues engineered stem cell-derived vessels comprising co-evolving endothelial cells and pericytes in a 3D hydrogel environment. With single-cell transcriptomes and a microfluidic migration assay, the authors characterized the transcriptional changes and cell communication signals between pericytes and endothelial cells during the process of neovascularization.