Core Viewpoint - The research presents a novel method for rapidly generating functional vascular organoids from induced pluripotent stem cells (iPSCs) through the orthogonal activation of transcription factors ETV2 and NKX3.1, demonstrating significant potential for applications in ischemia treatment and transplantation [3][11]. Group 1: Research Methodology - The study developed a simplified method to generate vascular organoids (VO) by using doxycycline-inducible or modRNA regulatory systems to activate transcription factors ETV2 and NKX3.1 [8]. - This method allows for the efficient co-differentiation of induced endothelial cells (iEC) and induced mural cells (iMC), producing functional 3D vascular organoids within 5 days without the need for extracellular matrix (ECM) embedding [8]. - Single-cell RNA sequencing revealed vascular heterogeneity, indicating that the timing of transcription factor activation influences the identity and heterogeneity of vascular cells [8]. Group 2: Research Findings - The vascular organoids formed perfusable blood vessels when implanted in immunodeficient mice, promoting vascular regeneration in models of hindlimb ischemia and islet transplantation [10][11]. - The research established a rapid and versatile vascular organoid platform with broad potential for vascular modeling, disease research, and regenerative cell therapy [13]. Group 3: Visual Representation - The cover image of the study illustrates the dual differentiation of human pluripotent stem cells into two vascular lineages—endothelial cells and mural cells—symbolizing a balanced and dynamic system inspired by the concept of yin and yang [7].

Core Viewpoint - The article discusses the development of a new method for rapidly generating functional vascular organoids from induced pluripotent stem cells (iPSCs) through the orthogonal activation of transcription factors ETV2 and NKX3.1, which significantly improves the efficiency and potential applications of vascular organoids in research and clinical settings [4][12][14]. Group 1: Vascular Organoids Overview - Vascular organoids (VO) are important models in cardiovascular research, capable of simulating dynamic interactions between endothelial cells and vascular wall cells, and replicating organ-specific vascular microenvironments [2][6]. - Current differentiation protocols for vascular organoids face challenges such as high heterogeneity, long time requirements, dependency on matrix gels and growth factors, high costs, and low in vivo vascularization capabilities [2][4]. Group 2: Research Development - A collaborative research team from Peking University and Harvard Medical School published a study in Cell Stem Cell detailing a new method for generating vascular organoids [3]. - The new method allows for the generation of uniform vascular organoids within 5 days from iPSCs, establishing a controllable vascular lineage differentiation model [4][10]. Group 3: Methodology and Findings - The method employs orthogonal activation of transcription factors ETV2 and NKX3.1 to induce iPSCs into endothelial cells (iEC) and mural cells (iMC), respectively, without the need for extracellular matrix [10][12]. - The vascular organoids formed have a diameter of approximately 250 μm and can mature further when embedded in extracellular matrix, leading to larger and more structured vessels [10][11]. - Single-cell RNA sequencing revealed that the duration of transcription factor activation influences the identity and heterogeneity of vascular cells, allowing for the potential to create specific types of vascular networks on demand [10][12]. Group 4: Applications and Implications - The vascular organoids demonstrated the ability to form perfused vessels when implanted in immunodeficient mice, promoting vascular reconstruction in ischemia and transplantation models [11][12]. - This research establishes a new platform for rapid and versatile vascular organoid generation, enhancing the potential for applications in vascular modeling, disease research, and regenerative cell therapies [14].