Generation of human hepatocytes from extended pluripotent stem cells

Wang, Qiming; Sun, Dihua; Liang, Zhen; Wang, Junyi; Zhong, Xinxing; Lyu, Yulin; Cao, Junning; Zheng, Lin; Du, Yuanyuan; Miao, Zewei; Lu, Shichun; Li, Cheng; Xu, Jun; Shi, Yan; Deng, Hongkui

doi:10.1038/s41422-020-0293-x

Cited by 23 publications

(20 citation statements)

References 18 publications

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“…Increasing the in vivo repopulation efficiency is a challenge for stem cell-derived hepatocytes. The iHep engraftment efficiency, in this study as well as many other similar studies [14,[34][35][36], was much lower than that achieved by engrafting primary human hepatocytes [37,38]. The low engraftment rate of iHeps is likely attributed to their relative immaturity.…”

Section: Discussioncontrasting

confidence: 44%

CRISPR-targeted genome editing of human induced pluripotent stem cell-derived hepatocytes for the treatment of Wilson’s disease

et al. 2022

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Background & Aims Wilson’s disease (WD) is an autosomal recessive disorder of copper metabolism caused by loss-of-function mutations in ATP7B , which encodes a copper-transporting protein. It is characterized by excessive copper deposition in tissues, predominantly in the liver and brain. We sought to investigate whether gene-corrected patient-specific induced pluripotent stem cell (iPSC)-derived hepatocytes (iHeps) could serve as an autologous cell source for cellular transplantation therapy in WD. Methods We first compared the in vitro phenotype and cellular function of ATP7B before and after gene correction using CRISPR/Cas9 and single-stranded oligodeoxynucleotides (ssODNs) in iHeps (derived from patients with WD) which were homozygous for the ATP7B R778L mutation (ATP7B R778L/R778L ). Next, we evaluated the in vivo therapeutic potential of cellular transplantation of WD gene-corrected iHeps in an immunodeficient WD mouse model ( Atp7b -/- / Rag2 -/- / Il2rg -/- ; ARG). Results We successfully created iPSCs with heterozygous gene correction carrying 1 allele of the wild-type ATP7B gene (ATP7B WT/- ) using CRISPR/Cas9 and ssODNs. Compared with ATP7B R778L/R778L iHeps, gene-corrected ATP7B WT/- iHeps restored i n vitro ATP7B subcellular localization, its subcellular trafficking in response to copper overload and its copper exportation function. Moreover, in vivo cellular transplantation of ATP7B WT/- iHeps into ARG mice via intra-splenic injection significantly attenuated the hepatic manifestations of WD. Liver function improved and liver fibrosis decreased due to reductions in hepatic copper accumulation and consequently copper-induced hepatocyte toxicity. Conclusions Our findings demonstrate that gene-corrected patient-specific iPSC-derived iHeps can rescue the in vitro and in vivo disease phenotypes of WD. These proof-of-principle data suggest that iHeps derived from gene-corrected WD iPSCs have potential use as an autologous ex vivo cell source for in vivo therapy of WD as well as other inherited liver disorders. Lay summary Gene correction restored ATP7B function in hepatocytes derived from induced pluripotent stem cells that originated from a patient with Wilson’s disease. These gene-corrected hepatocytes are potential cell sources for autologous cell therapy in patients with Wilson’s disease.

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Section: Discussioncontrasting

confidence: 44%

CRISPR-targeted genome editing of human induced pluripotent stem cell-derived hepatocytes for the treatment of Wilson’s disease

et al. 2022

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“…Human EPS cells may be efficiently differentiated into functional hepatocytes (EPS-Heps) through adding a pretreatment step before hepatic differentiation. 64 Importantly, compared with hepatocytes derived from conventional human pluripotent stem cells, EPS-Heps transcriptionally more resemble PHHs. However, the RIs are low (~2%), limiting its application in transplants.…”

Section: Generation Of Hepatocyte-like Cellsmentioning

confidence: 99%

Regenerative medicine for the hepatobiliary system: A review

Huang

Miyamoto

Hidaka

et al. 2020

J Hepato Biliary Pancreat

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The science of regenerative medicine aims to generate and use stem cells, engineered tissues, and artificial organs to restore or establish normal function. 1 Tremendous multidisciplinary efforts dedicated to liver regenerative medicine aim at providing transplantable cells, microtissues, and bioengineered whole livers via tissue engineering and maintaining partial liver function via extracorporeal support. 2,3 Although the liver has high regenerative capacity, endogenous and exogenous stimuli can still result in permanent tissue damage and impairment of liver function. Patients with endstage liver disease or acute liver failure often undergo liver transplantation, which is considerably limited by the lack

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“…Due to the difficulty in culturing primary hepatocytes, alternative cell sources have been explored, such as the directed differentiation of embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) towards hepatocytes 13 – 16 . In this context, specification towards the hepatocyte lineage was achieved by employing factors to activate (or inhibit) specific signaling pathways at various time points during culture to mimic in vivo developmental stages 17 .…”

Section: Introductionmentioning

confidence: 99%

“…Embedded in 3D extracellular matrix, LGR5 + CBCs can be coaxed to self-renew and generate various intestinal cell types in the presence of growth factors such as R-spondin (RSPO), epidermal growth factor (EGF) and noggin (NOG) while spontaneously organizing into crypt-villus structures. Organoid technology has since enabled the in vitro self-renewal of various epithelial cell types, which can be indefinitely expanded while spontaneously self-organizing into 3D structures that resemble their native counterparts 16 . Huch and colleagues described the culture of liver organoids derived from intrahepatic cholangiocytes, i.e., the epithelial cells that form the bile ducts 20 , 21 .…”

Section: Introductionmentioning

confidence: 99%

Hepatocyte organoids and cell transplantation: What the future holds

2021

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Historically, primary hepatocytes have been difficult to expand or maintain in vitro. In this review, we will focus on recent advances in establishing hepatocyte organoids and their potential applications in regenerative medicine. First, we provide a background on the renewal of hepatocytes in the homeostatic as well as the injured liver. Next, we describe strategies for establishing primary hepatocyte organoids derived from either adult or fetal liver based on insights from signaling pathways regulating hepatocyte renewal in vivo. The characteristics of these organoids will be described herein. Notably, hepatocyte organoids can adopt either a proliferative or a metabolic state, depending on the culture conditions. Furthermore, the metabolic gene expression profile can be modulated based on the principles that govern liver zonation. Finally, we discuss the suitability of cell replacement therapy to treat different types of liver diseases and the current state of cell transplantation of in vitro-expanded hepatocytes in mouse models. In addition, we provide insights into how the regenerative microenvironment in the injured host liver may facilitate donor hepatocyte repopulation. In summary, transplantation of in vitro-expanded hepatocytes holds great potential for large-scale clinical application to treat liver diseases.

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Generation of human hepatocytes from extended pluripotent stem cells

Cited by 23 publications

References 18 publications

CRISPR-targeted genome editing of human induced pluripotent stem cell-derived hepatocytes for the treatment of Wilson’s disease

CRISPR-targeted genome editing of human induced pluripotent stem cell-derived hepatocytes for the treatment of Wilson’s disease

Regenerative medicine for the hepatobiliary system: A review

Hepatocyte organoids and cell transplantation: What the future holds

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