References
-
Sun, Z. et al. Hepatocyte transplantation: the progress and the challenges. Hepatol. Commun. https://doi.org/10.1097/HC9.0000000000000266 (2023).
-
Hu, W. & Lazar, M. A. Modelling metabolic diseases and drug response using stem cells and organoids. Nat. Rev. Endocrinol. 18, 744–759 (2022).
-
Grossman, M. et al. Successful ex vivo gene therapy directed to liver in a patient with familial hypercholesterolaemia. Nat. Genet. 6, 335–341 (1994).
-
Hickey, R. D. et al. Curative ex vivo liver-directed gene therapy in a pig model of hereditary tyrosinemia type 1. Sci. Transl. Med. 8, 349ra399 (2016).
-
VanLith, C. J. et al. Ex vivo hepatocyte reprograming promotes homology-directed DNA repair to correct metabolic disease in mice after transplantation. Hepatol. Commun. 3, 558–573 (2019).
-
Artegiani, B. et al. Fast and efficient generation of knock-in human organoids using homology-independent CRISPR–Cas9 precision genome editing. Nat., Cell. Biol. 22, 321–331 (2020).
-
Kim, T. W. et al. Biphasic activation of WNT signaling facilitates the derivation of midbrain dopamine neurons from hESCs for translational use. Cell Stem Cell 28, 343–355 e345 (2021).
-
Touboul, T. et al. Generation of functional hepatocytes from human embryonic stem cells under chemically defined conditions that recapitulate liver development. Hepatology 51, 1754–1765 (2010).
-
Baxter, M. et al. Phenotypic and functional analyses show stem cell-derived hepatocyte-like cells better mimic fetal rather than adult hepatocytes. J. Hepatol. 62, 581–589 (2015).
-
Huang, P. et al. Direct reprogramming of human fibroblasts to functional and expandable hepatocytes. Cell Stem Cell 14, 370–384 (2014).
-
Grandy, R., Tomaz, R. A. & Vallier, L. Modeling disease with human inducible pluripotent stem cells. Annu. Rev. Pathol. 14, 449–468 (2019).
-
Xiang, C. et al. Long-term functional maintenance of primary human hepatocytes in vitro. Science 364, 399–402 (2019).
-
Fu, G. B. et al. Expansion and differentiation of human hepatocyte-derived liver progenitor-like cells and their use for the study of hepatotropic pathogens. Cell Res. 29, 8–22 (2019).
-
Hu, H. et al. Long-term expansion of functional mouse and human hepatocytes as 3D organoids. Cell 175, 1591–1606 e1519 (2018).
-
Zhang, K. et al. In vitro expansion of primary human hepatocytes with efficient liver repopulation capacity. Cell Stem Cell 23, 806–819 e804 (2018).
-
Katsuda, T. et al. Generation of human hepatic progenitor cells with regenerative and metabolic capacities from primary hepatocytes. eLife https://doi.org/10.7554/eLife.47313 (2019).
-
Kim, Y. et al. Small molecule-mediated reprogramming of human hepatocytes into bipotent progenitor cells. J. Hepatol. 70, 97–107 (2019).
-
Wang, C. et al. Dedifferentiation-associated inflammatory factors of long-term expanded human hepatocytes exacerbate their elimination by macrophages during liver engraftment. Hepatology 76, 1690–1705 (2022).
-
Zhang, K. et al. Ex vivo factor VIII-modified proliferating human hepatocytes therapy for haemophilia A. Cell Prolif. 56, e13467 (2023).
-
Yuan, X. et al. Preclinical efficacy and safety of encapsulated proliferating human hepatocyte organoids in treating liver failure. Cell Stem Cell 31, 484–498 e485 (2024).
-
Zhang, K. et al. Efficient expansion and CRISPR–Cas9-mediated gene correction of patient-derived hepatocytes for treatment of inherited liver diseases. Cell Stem Cell https://doi.org/10.1016/j.stem.2024.04.022 (2024).
-
Nicolas, C. T. et al. Ex vivo cell therapy by ectopic hepatocyte transplantation treats the porcine tyrosinemia model of acute liver failure. Mol. Ther. Methods Clin. Dev. 18, 738–750 (2020).
-
Huch, M. et al. In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration. Nature 494, 247–250 (2013).
-
Huch, M. et al. Long-term culture of genome-stable bipotent stem cells from adult human liver. Cell 160, 299–312 (2015).
-
Yanger, K. et al. Robust cellular reprogramming occurs spontaneously during liver regeneration. Genes Dev. 27, 719–724 (2013).
-
Tarlow, B. D. et al. Bipotential adult liver progenitors are derived from chronically injured mature hepatocytes. Cell Stem Cell 15, 605–618 (2014).
-
Tanimizu, N., Nishikawa, Y., Ichinohe, N., Akiyama, H. & Mitaka, T. Sry HMG box protein 9-positive (Sox9+) epithelial cell adhesion molecule-negative (EpCAM−) biphenotypic cells derived from hepatocytes are involved in mouse liver regeneration. J. Biol. Chem. 289, 7589–7598 (2014).
-
Yuan, X., Sun, Z., Wu, J., Hui, L. & Zhang, L. Cell therapy for liver diseases: from hepatocyte transplantation to bioartificial livers. Curr. Opin. Biomed. Eng. 30, 100530 (2024).
-
Labanieh, L. et al. Enhanced safety and efficacy of protease-regulated CAR-T cell receptors. Cell 185, 1745–1763 e1722 (2022).
-
Peng, W. C. et al. Inflammatory cytokine TNFα promotes the long-term expansion of primary hepatocytes in 3D culture. Cell 175, 1607–1619 e1615 (2018).
-
Guo, R. et al. IL6 supports long-term expansion of hepatocytes in vitro. Nat. Commun. 13, 7345 (2022).
-
Cox, D. B., Platt, R. J. & Zhang, F. Therapeutic genome editing: prospects and challenges. Nat. Med. 21, 121–131 (2015).
-
Dunbar, C. E. et al. Gene therapy comes of age. Science https://doi.org/10.1126/science.aan4672 (2018).
-
Ma, C. et al. CD47 and PD-L1 overexpression in proliferating human hepatocytes attenuated immune responses and ameliorated acute liver injury in mice. Am. J. Transplant. 23, 1832–1844 (2023).
-
Gao, Y. et al. Distinct gene expression and epigenetic signatures in hepatocyte-like cells produced by different strategies from the same donor. Stem Cell Rep. 9, 1813–1824 (2017).
-
Akbari, S. et al. Robust, long-term culture of endoderm-derived hepatic organoids for disease modeling. Stem Cell Rep., 13, 627–641 (2019).
-
Feng, S. et al. Large-scale generation of functional and transplantable hepatocytes and cholangiocytes from human endoderm stem cells. Cell Rep. 33, 108455 (2020).
-
Ma, C. et al. Single cell Raman spectroscopy to identify different stages of proliferating human hepatocytes for cell therapy. Stem Cell Res. Ther. 12, 555 (2021).
-
Qiao, S. et al. Functional proliferating human hepatocytes: in vitro hepatocyte model for drug metabolism, excretion, and toxicity. Drug Metab. Dispos. 49, 305–313 (2021).
-
Fennema, E., Rivron, N., Rouwkema, J., van Blitterswijk, C. & de Boer, J. Spheroid culture as a tool for creating 3D complex tissues. Trends Biotechnol. 31, 108–115 (2013).
-
Marsee, A. et al. Building consensus on definition and nomenclature of hepatic, pancreatic, and biliary organoids. Cell Stem Cell 28, 816–832 (2021).
-
Yamaguchi, T. et al. Generation of functional human hepatocytes in vitro: current status and future prospects. Inflamm. Regen. 39, 13 (2019).
-
Meng, X., Liu, A., Phangthavong, O. & Sun, Y. A novel strategy for treating acute liver failure: encapsulated proliferating human hepatocyte organoids. Biomater. Transl. 5, 444–446 (2024).
-
Yu, L. et al. Manganese is a potent inducer of lysosomal activity that inhibits de novo HBV infection. PLoS Pathog. 21, e1012800 (2025).
-
Lee, S. M., Schelcher, C., Demmel, M., Hauner, M. & Thasler, W. E. Isolation of human hepatocytes by a two-step collagenase perfusion procedure. J. Vis. Exp. https://doi.org/10.3791/50615 (2013).
-
Knobeloch, D. et al. Human hepatocytes: isolation, culture, and quality procedures. Methods Mol. Biol. 806, 99–120 (2012).
-
Broutier, L. et al. Culture and establishment of self-renewing human and mouse adult liver and pancreas 3D organoids and their genetic manipulation. Nat. Protoc. 11, 1724–1743 (2016).
-
Sun, L. et al. Modelling liver cancer initiation with organoids derived from directly reprogrammed human hepatocytes. Nat. Cell. Biol. 21, 1015–1026 (2019).
-
Ogawa, S. et al. Three-dimensional culture and cAMP signaling promote the maturation of human pluripotent stem cell-derived hepatocytes. Development 140, 3285–3296 (2013).
-
Azuma, H. et al. Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice. Nat. Biotechnol. 25, 903–910 (2007).
