A two-step lineage reprogramming strategy to generate functionally competent human hepatocytes from fibroblasts

Bingqing Xie,Weifeng Lai,Ying Pu ,Huangfan Xie,Xiaonan Zhang,Min Wu,Xuehui Lyu,Jun Xu,Dihua Sun ,Cheng Li,Shichun Lu,Shicheng Sun,Hui Shen,Zhenghong Yuan,Shiyu Li,Yan Shi,Chengang Xiang,Qiming Wang,Guang‐Ya Li ,Jinping Jia ,Peng Du,Yifang Liu,Yuanyuan Du,Kuanhui Xiang,Sitong Chen,Hongkui Deng

doi:10.1038/s41422-019-0196-x

Bingqing Xie, Weifeng Lai + Show 24 more

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https://doi.org/10.1038/s41422-019-0196-x

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Terminally differentiated cells can be generated by lineage reprogramming, which is, however, hindered by incomplete conversion with residual initial cell identity and partial functionality. Here, we demonstrate a new reprogramming strategy by mimicking the natural regeneration route, which permits generating expandable hepatic progenitor cells and functionally competent human hepatocytes. Fibroblasts were first induced into human hepatic progenitor-like cells (hHPLCs), which could robustly expand in vitro and efficiently engraft in vivo. Moreover, hHPLCs could be efficiently induced into mature human hepatocytes (hiHeps) in vitro, whose molecular identity highly resembles primary human hepatocytes (PHHs). Most importantly, hiHeps could be generated in large quantity and were functionally competent to replace PHHs for drug-metabolism estimation, toxicity prediction and hepatitis B virus infection modeling. Our results highlight the advantages of the progenitor stage for successful lineage reprogramming. This strategy is promising for generating other mature human cell types by lineage reprogramming.

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The generation of desired functional cells is a long-standing goal of stem cell research and regenerative medicine
Generation of proliferative human hepatic progenitor cells To generate human hepatic progenitor cells from human embryonic fibroblasts (HEFs), we selected a pool of transcription factors (TFs) as candidates based on (1) their importance in hepatic organogenesis, and (2) computational analysis of the highly expressed TFs in human fetal liver cells, a specified type of human hepatic progenitor cells that gives rise to hepatocytes in adult liver (Supplementary information, Table S1)
Systematic characterization revealed that induced hepatocytes recapitulated the mature characteristics of primary human hepatocytes, making it feasible to utilize these cells in a wide range of applications, such as drug discovery and liver disease modeling

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Introduction

The generation of desired functional cells is a long-standing goal of stem cell research and regenerative medicine. In the past few decades, extensive efforts have been made to develop various strategies to produce functional cells by differentiation from pluripotent stem cells or conversion from a distinct cell type by lineage reprogramming.[1,2] for many functional lineages, these strategies encountered a major challenge: the cells generated in vitro rarely obtained mature cell identity and functionality.[3,4] most of these cells retained the expression of fetal markers and/or were lacking in the expression of adult markers, leading to an immature cell state.[5,6] it remains a challenge to develop a new approach to generate desired cells in vitro that resemble their counterparts isolated from the native tissue. A new strategy to overcome these barriers would greatly improve the functionality of cells generated by lineage reprogramming

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