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Engineering perfusable vascular networks in organoids-on-chip

ABSTRACT: Vascular networks are critical for the development and maintenance of human tissues, as they support metabolism and regulate tissue growth and function. In vitro models such as organoids and organs-on-chip often have a limited capacity to recapitulate in vivo functional vascular networks and their integration within tissues. Most existing systems fail to mimic the structural and functional complexity of native capillary beds, lack physiological flow dynamics, and do not support vascular circulation. Here, we present a fully stem cell-derived microfluidic platform capable of generating perfusable vascular network organoids-on-chip with capillary-scale vessels, endothelial responses to biophysical forces, and physiologically accurate gene expression. Using this platform, we generate vascularized heart organoids-on-chip that replicate the dense capillary architecture of the heart and exhibit organ-specific vascular features and gene expression. These results establish a scalable and physiologically relevant approach for engineering and studying vascularized organoids under dynamic flow, with broad applications in developmental biology, disease modeling, and multi-organ in vitro systems.

ORGANISM(S): Homo sapiens

PROVIDER: GSE295488 | GEO | 2026/04/22

REPOSITORIES: GEO

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Project description:Primitive macrophages enhance a perivascular niche to enable vascular perfusion in human cardiac tissue The intricate anatomical structure and high cellular density of the myocardium significantly complicate the bioengineering of vascular networks within cardiac tissues, creating challenges in establishing a perfusable and stable vasculature within these tissues. Emerging evidence from murine in vivo studies underscores the significant role of resident cardiac macrophages in facilitating cardiac regeneration post-injury, specifically their role in enhancing angiogenesis. Here, for the first time, we integrate human pluripotent stem cell derived macrophages, resembling primitive yolk-sac derived macrophages, within human in vitro vascularized heart-on-chip platforms. The incorporation of primitive macrophages had a profound impact on the long term functionality of microvascularized cardiac tissue, particularly in enabling formation of perfusable vasculature with a stable barrier function that persisted for two weeks in culture. These effects were contingent on physical cell-cell interactions and significantly diminished in transwell culture. The inclusion of primitive macrophages mitigated tissue cytotoxicity and curtailed the release of cell-free mitochondrial-DNA, underscoring their indispensable role for mitigating cell damage in bioengineered tissues. Furthermore, their incorporation upregulated the secretion of pro-angiogenic, matrix remodeling and cardioprotective cytokines such as MMP-12, MMP-2, Angiopoietin-like 1, NRG-3, SIGIRR and Adiponectin. RNA sequencing disclosed upregulation of cardiac maturation (TTNI3, SCN5A, MYL2, and RYR2), and endothelial cells genes (PDGF-B, PECAM-1 and CDH5) as well as enhancement of angiogenic signaling, Wnt and PDGF pathways. Collectively, our results offer valuable insights into the integral role of primitive macrophages in directing long-term functional vascularization of cardiac tissues, paving the way for novel therapeutic strategies and advancing heart-on-a-chip systems.

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Engineering perfusable vascular networks in organoids-on-chip

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