During early liver organogenesis, newly specified hepatic cells delaminatefromtheforegut endodermal sheet andform aliver bud8, a condensedtissue mass that is soon vascularized. Such large-scale morphogeneticchanges depend on the exquisite orchestration of signals between endodermalepithelial, mesenchymal and endothelial progenitors before bloodperfusion5. These observations led us to propose that three-dimensionalliver-bud formation can be recapitulated in vitro by culturing hepaticendoderm cells with endothelial and mesenchymal lineages (Fig. 1a). Toexamine this hypothesis, we first prepared hepatic endoderm cells fromhuman iPSCs (iPSC-HEs) by directed differentiation, producing approximately80% of the treated cells expressed the hepatic marker HNF4A,which is involved in cell fate determination
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