Self-Organization of Sinusoidal Vessels in Pluripotent Stem Cell-derived Human Liver Bud Organoids.
Norikazu SaikiYasunori NioYosuke YoneyamaShuntaro KawamuraKentaro IwasawaEri KawakamiKohei ArakiJunko FukumuraTsuyoshi SakairiTamaki KonoRio OhmuraMasaru KoidoMasaaki FunataWendy L ThompsonPamela Cruz-EncarnacionYa-Wen ChenTakanori TakebePublished in: bioRxiv : the preprint server for biology (2024)
The induction of tissue-specific vessels in in vitro living tissue systems remains challenging. Here, we directly differentiated human pluripotent stem cells into CD32b + putative liver sinusoidal progenitors (iLSEP) by dictating developmental pathways. By devising an inverted multilayered air-liquid interface (IMALI) culture, hepatic endoderm, septum mesenchyme, arterial and sinusoidal quadruple progenitors self-organized to generate and sustain hepatocyte-like cells neighbored by divergent endothelial subsets composed of CD32b low CD31 high , LYVE1 + STAB1 + CD32b high CD31 low THBD - vWF - , and LYVE1 - THBD + vWF + cells. Wnt2 mediated sinusoidal-to-hepatic intercellular crosstalk potentiates hepatocyte differentiation and branched endothelial network formation. Intravital imaging revealed iLSEP developed fully patent human vessels with functional sinusoid-like features. Organoid-derived hepatocyte- and sinusoid-derived coagulation factors enabled correction of in vitro clotting time with Factor V, VIII, IX, and XI deficient patients' plasma and rescued the severe bleeding phenotype in hemophilia A mice upon transplantation. Advanced organoid vascularization technology allows for interrogating key insights governing organ-specific vessel development, paving the way for coagulation disorder therapeutics.
Keyphrases
- endothelial cells
- pluripotent stem cells
- nk cells
- newly diagnosed
- end stage renal disease
- high resolution
- induced apoptosis
- chronic kidney disease
- liver injury
- metabolic syndrome
- atrial fibrillation
- single cell
- photodynamic therapy
- signaling pathway
- mesenchymal stem cells
- mass spectrometry
- oxidative stress
- drug induced
- insulin resistance