Dedifferentiation‐associated inflammatory factors of long‐term expanded human hepatocytes exacerbate their elimination by macrophages during liver engraftment

Abstract Background and Aims Hepatocyte transplantation has been demonstrated to be effective to treat liver metabolic disease and acute liver failure. Nevertheless, the shortage of donor hepatocytes restrained its application in clinics. To expand human hepatocytes at a large scale, several dedifferentiation‐based protocols have been established, including proliferating human hepatocytes (ProliHH). However, the decreased transplantation efficiency of these cells after long‐term expansion largely impedes their application. Approach and Results We found that accompanied with dedifferentiation, long‐term cultured ProliHH (lc‐ProliHH) up‐regulated a panel of chemokines and cytokines related to innate immunity, which were referred to as dedifferentiation‐associated inflammatory factors (DAIF). DAIF elicited excessive macrophage responses, accounting for the elimination of lc‐ProliHH specifically during engraftment. Two possible strategies to increase ProliHH transplantation were then characterized. Blockage of innate immune response by dexamethasone reverted the engraftment and repopulation of lc‐ProliHH to a level comparable to primary hepatocytes, resulting in improved liver function and a better survival of fumarylacetoacetate hydrolase–deficient mice. Alternatively, rematuration of lc‐ProliHH as organoids reduced the expression of DAIF and led to markedly improved engraftment. Conclusions These results revealed that lc‐ProliHH triggers exacerbated macrophage activation by DAIF and provided potential solutions for clinical transplantation of lc‐ProliHH.