Blood vessels are vital to nearly every tissue in the body, delivering oxygen and nutrients, maintaining hemostasis, and modulating inflammation. Recreating these functional vascular networks in vitro is foundational for both basic research and translational applications. Yet most methods for generating vascular organoids (VOs) from stem cells remain slow, inefficient, and often lack the complexity needed for therapeutic relevance—until now.
In a new study published in Cell Stem Cell, titled “Rapid generation of functional vascular organoids via simultaneous transcription factor activation of endothelial and mural lineages,” researchers from Boston Children’s Hospital, Harvard Medical School, and others present a streamlined method for building VOs from induced pluripotent stem cells (iPSCs). The approach accelerates the formation of 3D microvascular networks while maintaining critical cell–cell interactions and functional performance in vivo.
A key challenge in vascular organoid generation has been the coordinated development of endothelial cells (ECs) and mural cells (MCs)—two lineages that must differentiate in tandem to form stable vessels. These cell types rely on tightly linked signaling pathways (including PDGFB, Notch, and TGFβ), yet require distinct and often incompatible culture conditions, making co-differentiation difficult.
To overcome this, the team employed orthogonal activation of two transcription factors: ETV2 and NKX3.1. Using either doxycycline (Dox)-inducible or modified mRNA (modRNA) systems, they drove the independent yet synchronized differentiation of both lineages in a suspension culture. Within five days—and prior to extracellular matrix (ECM) embedding—the cells self-assembled into primitive vascular networks. Upon ECM exposure, the organoids matured further, forming larger and more structured vessels.
Single-cell RNA sequencing confirmed vascular heterogeneity and revealed that timing the expression of transcription factors enabled modulation of angiogenic and arterial endothelial phenotypes. When transplanted into immunodeficient mice, the organoids connected with host vasculature and supported perfusion. In two disease models—hindlimb ischemia and pancreatic islet transplantation—the vascular organoids promoted robust revascularization.
The study also acknowledges limitations. Because both transcription factors are induced using the same Dox trigger, independent control of EC and MC development remains constrained. Future refinements using orthogonal induction systems (such as combining Dox and modRNA) could allow more flexible modulation. The current suspension culture also lacks perfusion, limiting the ability to simulate shear stress, a critical regulator of endothelial maturation. Incorporating VOs into perfusable platforms may enhance physiological relevance. Additionally, whether iPSC-derived ECs can acquire tissue-specific identity post-engraftment remains to be investigated.
Still, the method represents a significant advance. By rapidly producing self-assembling, functional vascular organoids with controlled dual-lineage specification, the system offers a scalable platform for modeling vascular disease, testing therapeutics, and engineering tissues. It sets a new bar for speed and sophistication in vascular organoid research—and holds promise for future regenerative medicine applications.
The post Rapid Formation of Vascular Organoids from iPSCs via TF Activation appeared first on GEN - Genetic Engineering and Biotechnology News.
In a new study published in Cell Stem Cell, titled “Rapid generation of functional vascular organoids via simultaneous transcription factor activation of endothelial and mural lineages,” researchers from Boston Children’s Hospital, Harvard Medical School, and others present a streamlined method for building VOs from induced pluripotent stem cells (iPSCs). The approach accelerates the formation of 3D microvascular networks while maintaining critical cell–cell interactions and functional performance in vivo.
A key challenge in vascular organoid generation has been the coordinated development of endothelial cells (ECs) and mural cells (MCs)—two lineages that must differentiate in tandem to form stable vessels. These cell types rely on tightly linked signaling pathways (including PDGFB, Notch, and TGFβ), yet require distinct and often incompatible culture conditions, making co-differentiation difficult.
Dual lineage transcription activation
To overcome this, the team employed orthogonal activation of two transcription factors: ETV2 and NKX3.1. Using either doxycycline (Dox)-inducible or modified mRNA (modRNA) systems, they drove the independent yet synchronized differentiation of both lineages in a suspension culture. Within five days—and prior to extracellular matrix (ECM) embedding—the cells self-assembled into primitive vascular networks. Upon ECM exposure, the organoids matured further, forming larger and more structured vessels.
Single-cell RNA sequencing confirmed vascular heterogeneity and revealed that timing the expression of transcription factors enabled modulation of angiogenic and arterial endothelial phenotypes. When transplanted into immunodeficient mice, the organoids connected with host vasculature and supported perfusion. In two disease models—hindlimb ischemia and pancreatic islet transplantation—the vascular organoids promoted robust revascularization.
Limitations and potential therapeutic applications
The study also acknowledges limitations. Because both transcription factors are induced using the same Dox trigger, independent control of EC and MC development remains constrained. Future refinements using orthogonal induction systems (such as combining Dox and modRNA) could allow more flexible modulation. The current suspension culture also lacks perfusion, limiting the ability to simulate shear stress, a critical regulator of endothelial maturation. Incorporating VOs into perfusable platforms may enhance physiological relevance. Additionally, whether iPSC-derived ECs can acquire tissue-specific identity post-engraftment remains to be investigated.
Still, the method represents a significant advance. By rapidly producing self-assembling, functional vascular organoids with controlled dual-lineage specification, the system offers a scalable platform for modeling vascular disease, testing therapeutics, and engineering tissues. It sets a new bar for speed and sophistication in vascular organoid research—and holds promise for future regenerative medicine applications.
The post Rapid Formation of Vascular Organoids from iPSCs via TF Activation appeared first on GEN - Genetic Engineering and Biotechnology News.