Abstract
Integration of resident immune cells into in vitro organoid models is important for accurately recapitulating native tissue physiology. Here, we present a protocol for integrating liver-resident macrophages, Kupffer cells, into liver organoid models derived from human induced pluripotent stem cells (iPSCs). We describe procedures for generating Kupffer cell progenitors and hepatic endoderm from iPSCs, followed by detailed steps for establishing liver organoids containing Kupffer cells (KuLOs). This protocol provides a platform for investigating the roles of Kupffer cells in liver development and diseases. For complete details on the use and execution of this protocol, please refer to Li et al.1.