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

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

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