Integration of Kupffer cells into human iPSC-derived liver organoids for modeling liver dysfunction in sepsis.

Maximizing the potential of human liver organoids (LOs) for modeling human septic liver requires the integration of innate immune cells, particularly resident macrophage Kupffer cells. In this study, we present a strategy to generate LOs containing Kupffer cells (KuLOs) by recapitulating fetal liver hematopoiesis using human induced pluripotent stem cell (hiPSC)-derived erythro-myeloid progenitors (EMPs), the origin of tissue-resident macrophages, and hiPSC-derived LOs. Remarkably, LOs actively promote EMP hematopoiesis toward myeloid and erythroid lineages. Moreover, supplementing with macrophage colony-stimulating factor (M-CSF) proves crucial in sustaining the hematopoietic population during the establishment of KuLOs. Exposing KuLOs to sepsis-like endotoxins leads to significant organoid dysfunction that closely resembles the pathological characteristics of the human septic liver. Furthermore, we observe a notable functional recovery in KuLOs upon endotoxin elimination, which is accelerated by using Toll-like receptor-4-directed endotoxin antagonist. Our study represents a comprehensive framework for integrating hematopoietic cells into organoids, facilitating in-depth investigations into inflammation-mediated liver pathologies. [Display omitted] • hiPSC-derived EMPs undergo hematopoiesis in a liver organoid environment • M-CSF is essential to establish liver organoids containing Kupffer cells (KuLOs) • LPS/IFN-γ-stimulated KuLOs resemble sepsis-associated liver dysfunction • Septic KuLOs have self-recovery ability Li et al. report the generation of liver organoids containing Kupffer cells (KuLOs) by recapitulating erythro-myeloid progenitor (EMP) hematopoiesis during liver development. LPS/IFN-γ-treated KuLOs can mimic the pathological events of the human septic liver. The impaired organoid functions can recover under stimulus-free conditions. [ABSTRACT FROM AUTHOR]