Human induced pluripotent stem cell-derived kidney organoids have potential for disease modeling and to be developed into clinically transplantable auxiliary tissue. However, they lack a functional vasculature, and the sparse endogenous endothelial cells (ECs) are lost upon prolonged culture in vitro, limiting maturation and applicability. Here, we use intracoelomic transplantation in chicken embryos followed by single-cell RNA sequencing and advanced imaging platforms to induce and study vasculogenesis in kidney organoids. We show expansion of human organoid-derived ECs that reorganize into perfused capillaries and form a chimeric vascular network with host-derived blood vessels. Ligand-receptor analysis infers extensive potential interactions of human ECs with perivascular cells upon transplantation, enabling vessel wall stabilization. Perfused glomeruli display maturation and morphogenesis to capillary loop stage. Our findings demonstrate the beneficial effect of vascularization on not only epithelial cell types, but also the mesenchymal compartment, inducing the expansion of ´on target´ perivascular stromal cells, which in turn are required for further maturation and stabilization of the neo-vasculature. The here described vasculogenic capacity of kidney organoids will have to be deployed to achieve meaningful glomerular maturation and kidney morphogenesis in vitro.
We are grateful to Christian Freund (hiPSC core facility, LUMC, Leiden, the Netherlands) for providing hiPSC lines (LUMC0072 and LUMC0020), and Melissa Little (Murdoch Children’s Research Institute, Melbourne, Australia) for iPSC-MAFB. We acknowledge the support of Saskia van der Wal-Maas (Department of Anatomy & Embryology, LUMC, Leiden, the Netherlands), Conny van Munsteren (Department of Anatomy & Embryology, LUMC, Leiden, the Netherlands), Manon Zuurmond (LUMC, Leiden, the Netherlands), George Galaris (LUMC, Leiden, the Netherlands), and Annemarie de Graaf (LUMC, Leiden, the Netherlands). This work is supported by the partners of Regenerative Medicine Crossing Borders (RegMedXB) and Health Holland, Top Sector Life Sciences & Health. M.Koning is supported by ‘Nephrosearch Stichting tot steun van het wetenschappelijk onderzoek van de afdeling Nierziekten van het LUMC’. S.J. Dumas is supported by a Marie Skłodowska-Curie fellowship (grant agreement No 846615) from the European Union’s Horizon 2020 research and innovation program. M. Borri is supported by the ‘Fonds voor Wetenschappelijk Onderzoek’ (FWO). L. Lin is supported by the DFF Sapere Aude Starting grant (8048-00072A). Y. Luo is supported by the Danish Research Council (9041-00317B) and European Union’s Horizon 2020 research and innovation program under grant agreement No 899417. H.S. Spijker is supported by a Kolff grant from the Dutch Kidney Foundation. P. Carmeliet is supported by Grants from Methusalem funding (Flemish government), the Fund for Scientific Research-Flanders (FWO-Vlaanderen), the European Research Council ERC Advanced Research Grant EU- ERC74307, and NNF Laureate Research Grant from Novo Nordisk Foundation (Denmark). C.W. van den Berg is supported by the Wiyadharma fellowship (Bontius stichting-LUMC). C.W. van den Berg and T.J. Rabelink are supported by The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), The Novo Nordisk Foundation Center for Stem Cell Medicine is supported by Novo Nordisk Foundation grants (NNF21CC0073729).
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