Hemangio-organoids derived from human pluripotent stem cells support the formation of dendritic cells

Background Organoid systems have emerged as powerful models to recapitulate organ development and function; however, the developmental potential and applications of hemangio-organoids remain poorly understood. Given the increasing interest in organoid-based transplantation strategies, the development of hemangio-organoids capable of generating functional hematopoietic cells may provide a promising platform for hematopoietic and immune cell therapies. Methods Human pluripotent stem cells (hPSCs) were cultured in a three-dimensional suspension system to generate hPSC-derived spheres (HPSpheres) and induced to differentiate into hemangio-organoids. Hemato-endothelial potential and endothelial-to-hematopoietic transition (EHT) were assessed, followed by differentiation into hematopoietic progenitor cells (HPCs) and myeloid progenitors. Dendritic cell differentiation was evaluated using gene expression analysis of key transcription factors (PU.1, IRF8, IRF4, and BATF3) and flow cytometric analysis of dendritic cell surface markers (CD80, CD86, CD83, and CD141). Functional capacity was assessed by measuring the ability of hemangio-organoid–derived mature dendritic cells to stimulate interferon-gamma (IFN-γ)–secreting T cells. Differentiation was achieved using a stepwise protocol based on small molecules and growth factors without genetic manipulation. Results Hemangio-organoids derived from hPSCs successfully recapitulated early hematopoietic development and underwent EHT to generate HPCs. These progenitors differentiated into myeloid progenitors and subsequently into mature dendritic cells, as evidenced by the expression of dendritic cell–associated transcription factors and surface markers. The resulting mature dendritic cells expressed the CD141 receptor and demonstrated functional activity by inducing proliferation of IFN-γ–secreting T cells. The stepwise, non-genetic differentiation protocol proved to be reliable, reproducible, scalable, and more cost-effective than previously reported approaches. Conclusions This study demonstrates that hPSC-derived hemangio-organoids can serve as an efficient platform for generating functional dendritic cells through a defined and scalable differentiation protocol. These findings highlight the potential of hemangio-organoids as a source of clinically relevant immune cells and support their future application in immunotherapy and regenerative medicine.