Generation of integration-free induced hepatocyte-like cells from mouse fibroblasts

Jonghun Kim; Kee-Pyo Kim; Kyung Tae Lim; Seung Chan Lee; Juyong Yoon; Guangqi Song; Seon In Hwang; Hans R Schöler; Tobias Cantz; Dong Wook Han

doi:10.1038/srep15706

Generation of integration-free induced hepatocyte-like cells from mouse fibroblasts

Sci Rep. 2015 Oct 27:5:15706. doi: 10.1038/srep15706.

Authors

Jonghun Kim¹, Kee-Pyo Kim², Kyung Tae Lim¹, Seung Chan Lee¹, Juyong Yoon², Guangqi Song³, Seon In Hwang¹, Hans R Schöler^{2

4}, Tobias Cantz^{2

3}, Dong Wook Han^{1

5}

Affiliations

¹ Department of Stem Cell Biology, School of Medicine, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.
² Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Röntgenstrasse 20, 48149 Münster, Germany.
³ REBIRTH Cluster of Excellence, Hannover Medical School, Hannover 30625, Germany.
⁴ University of Münster, Medical Faculty, Domagkstrasse 3, 48149 Münster, Germany.
⁵ KU Open-Innovation Center, Institute of Biomedical Science &Technology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, Republic of Korea.

Abstract

The ability to generate integration-free induced hepatocyte-like cells (iHeps) from somatic fibroblasts has the potential to advance their clinical application. Here, we have generated integration-free, functional, and expandable iHeps from mouse somatic fibroblasts. To elicit this direct conversion, we took advantage of an oriP/EBNA1-based episomal system to deliver a set of transcription factors, Gata4, Hnf1a, and Foxa3, to the fibroblasts. The established iHeps exhibit similar morphology, marker expression, and functional properties to primary hepatocytes. Furthermore, integration-free iHeps prolong the survival of fumarylacetoacetate-hydrolase-deficient (Fah(-/-)) mice after cell transplantation. Our study provides a novel concept for generating functional and expandable iHeps using a non-viral, non-integrating, plasmid-based system that could facilitate their pharmaceutical and biomedical application.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cadherins / genetics
Cadherins / metabolism
Cell Differentiation
Cells, Cultured
Cytochrome P-450 Enzyme System / genetics
Cytochrome P-450 Enzyme System / metabolism
Embryo, Mammalian / cytology
Fibroblasts / cytology*
Fibroblasts / metabolism
GATA4 Transcription Factor / genetics*
Hepatocyte Nuclear Factor 1-alpha / genetics*
Hepatocyte Nuclear Factor 3-gamma / genetics*
Hepatocytes / cytology*
Hepatocytes / metabolism
Hydrolases / deficiency
Hydrolases / genetics
Karyotyping
Keratin-18 / genetics
Keratin-18 / metabolism
Mice
Mice, Inbred C57BL
Mice, Knockout
Microscopy, Fluorescence
Serum Albumin / genetics
Serum Albumin / metabolism
Zonula Occludens-1 Protein / genetics
Zonula Occludens-1 Protein / metabolism

Substances

Cadherins
GATA4 Transcription Factor
Hepatocyte Nuclear Factor 1-alpha
Keratin-18
Serum Albumin
Tjp1 protein, mouse
Zonula Occludens-1 Protein
Hepatocyte Nuclear Factor 3-gamma
Cytochrome P-450 Enzyme System
Hydrolases
fumarylacetoacetase