Highlights
- •Human hepatic progenitors (hCdHs) are generated from adult hepatocytes.
- •HGF is required for chemical reprogramming induced by A83-01 and CHIR99021.
- •hCdHs proliferate for at least 10 passages without losing differentiation potential in vitro.
- •Bipotent hCdHs can repopulate injured liver and acquire functional properties.
Background & Aims
Currently, much effort is directed towards the development of new cell sources for
clinical therapy using cell fate conversion by small molecules. Direct lineage reprogramming
to a progenitor state has been reported in terminally differentiated rodent hepatocytes,
yet remains a challenge in human hepatocytes.
Methods
Human hepatocytes were isolated from healthy and diseased donor livers and reprogrammed
into progenitor cells by 2 small molecules, A83-01 and CHIR99021 (AC), in the presence
of EGF and HGF. The stemness properties of human chemically derived hepatic progenitors
(hCdHs) were tested by standard in vitro and in vivo assays and transcriptome profiling.
Results
We developed a robust culture system for generating hCdHs with therapeutic potential.
The use of HGF proved to be an essential determinant of the fate conversion process.
Based on functional evidence, activation of the HGF/MET signal transduction system
collaborated with A83-01 and CHIR99021 to allow a rapid expansion of progenitor cells
through the activation of the ERK pathway. hCdHs expressed hepatic progenitor markers
and could self-renew for at least 10 passages while retaining a normal karyotype and
potential to differentiate into functional hepatocytes and biliary epithelial cells
in vitro. Gene expression profiling using RNAseq confirmed the transcriptional reprogramming
of hCdHs towards a progenitor state and the suppression of mature hepatocyte transcripts.
Upon intrasplenic transplantation in several models of therapeutic liver repopulation,
hCdHs effectively repopulated the damaged parenchyma.
Conclusion
Our study is the first report of successful reprogramming of human hepatocytes to
a population of proliferating bipotent cells with regenerative potential. hCdHs may
provide a novel tool that permits expansion and genetic manipulation of patient-specific
progenitors to study regeneration and the repair of diseased livers.
Lay summary
Human primary hepatocytes were reprogrammed towards hepatic progenitor cells by a
combined treatment with 2 small molecules, A83-01 and CHIR99021, and HGF. Chemically
derived hepatic progenitors exhibited a high proliferation potential and the ability
to differentiate into hepatocytes and biliary epithelial cells both in vitro and in vivo. This approach enables the generation of patient-specific hepatic progenitors and
provides a platform for personal and stem cell-based regenerative medicine.
Graphical abstract
Graphical Abstract
Keywords
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Article info
Publication history
Published online: September 18, 2018
Accepted:
September 10,
2018
Received in revised form:
August 2,
2018
Received:
October 20,
2017
Identification
Copyright
© 2018 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
