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bioRxiv · 10.1101/2024.07.02.601804

Self-Organization of Sinusoidal Vessels in Pluripotent Stem Cell-derived Human Liver Bud Organoids

Saiki, N.·Nio, Y.·Yoneyama, Y.·Kawamura, S.·Iwasawa, K.·Kawakami, E.·Araki, K.·Fukumura, J.·Sakairi, T.·Ohmura, R.·Koido, M.·Funata, M.·Thompson, W. L.·Cruz-Encarnacion, P.·Chen, Y.-W.·Takebe, T.
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Abstract

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 CD32blowCD31high, LYVE1+STAB1+CD32bhighCD31lowTHBD-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.

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Saiki, N., Nio, Y., Yoneyama, Y., Kawamura, S., Iwasawa, K., Kawakami, E., Araki, K., Fukumura, J., Sakairi, T., Ohmura, R., Koido, M., Funata, M., Thompson, W. L., Cruz-Encarnacion, P., Chen, Y.-W., Takebe, T.. 2024-07-04. Self-Organization of Sinusoidal Vessels in Pluripotent Stem Cell-derived Human Liver Bud Organoids. https://doi.org/10.1101/2024.07.02.601804

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