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Liver stem cells are dormant and generally not detectable in the healthy adult liver. However, injury triggers the activation of an Lgr5⁺ epithelial progenitor cell population. Single Lgr5⁺ cells from adult mouse liver can be isolated and expanded into a 3D culture system to generate hepatic organoids that possess clonogenic potential and can be maintained long-term. Use to establish and maintain hepatic organoids from mouse ductal fragments. For human cultures, Epcam⁺ bile duct-derived bipotent progenitors can be isolated for expansion into human hepatic organoids. Use to generate mature hepatic organoids from fresh or frozen human liver tissue.