Robust expansion and functional maturation of human hepatoblasts by chemical strategy

Tingcai Pan,Yin-Xiong Li,Kai You,Ning Wang,Ji Fang,Yan Chen,Jiaye Zhang,Jiawang Tao,Yuanqi Zhuang,Yi Gao,Yingying Xu,Fan Yang,Xianhua Lin,Shenglin Tan,Anteneh Getachew

doi:10.1186/s13287-021-02233-9

Abstract

BackgroundChemically strategies to generate hepatic cells from human pluripotent stem cells (hPSCs) for the potential clinical application have been improved. However, producing high quality and large quantities of hepatic cells remain challenging, especially in terms of step-wise efficacy and cost-effective production requires more improvements.MethodsHere, we systematically evaluated chemical compounds for hepatoblast (HB) expansion and maturation to establish a robust, cost-effective, and reproducible methodology for self-renewal HBs and functional hepatocyte-like cell (HLC) production.ResultsThe established chemical cocktail could enable HBs to proliferate nearly 3000 folds within 3 weeks with preserved bipotency. Moreover, those expanded HBs could be further efficiently differentiated into homogenous HLCs which displayed typical morphologic features and functionality as mature hepatocytes including hepatocyte identity marker expression and key functional activities such as cytochrome P450 metabolism activities and urea secretion. Importantly, the transplanted HBs in the injured liver of immune-defect mice differentiated as hepatocytes, engraft, and repopulate in the injured loci of the recipient liver.ConclusionTogether, this chemical compound-based HLC generation method presents an efficient and cost-effective platform for the large-scale production of functional human hepatic cells for cell-based therapy and drug discovery application.

Highlights

Strategies to generate hepatic cells from human pluripotent stem cells for the potential clinical application have been improved
HBs are capable of self-renewal and have the potency to differentiate into hepatocytes and expand purified HBs derived from human pluripotent stem cells (hPSCs) which would be an ideal strategy to enable massive hepatic cell production
We evaluated cytochrome P450 (CYP) enzymes’ metabolic activities in derived hepatocyte-like cells (HLCs), as metabolism and detoxification in the liver are mainly executed by these enzymes in hepatocytes

Summary

Introduction

Strategies to generate hepatic cells from human pluripotent stem cells (hPSCs) for the potential clinical application have been improved. Producing high quality and large quantities of hepatic cells remain challenging, especially in terms of step-wise efficacy and cost-effective production requires more improvements. A robust and cost-effective expansion and differentiation method for large-scale production of functional human hepatocytes could be beneficial for many potential clinical applications, including hepatic cell transplantation, bio-artificial liver, drug development, and disease modeling [1, 2]. Recent studies reported hepatic expansion and maturation methods mostly applied expensive growth factors, which leads to critical challenges such as the high cost and protocol reproducibility in large-scale production [7,8,9]. This suggested a costeffective and reproducible approach by using chemical compounds for producing scalable and integrated functional hepatocytes

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Discussion

Conclusion