Oct4 and Hnf4α-induced hepatic stem cells ameliorate chronic liver injury in liver fibrosis model

PLoS One. 2019 Aug 12;14(8):e0221085. doi: 10.1371/journal.pone.0221085. eCollection 2019.

Abstract

Direct conversion from fibroblasts to generate hepatocyte like-cells (iHeps) bypassing the pluripotent state has been described in previous reports as an attractive method acquiring hepatocytes for cell-based therapy. The limited proliferation of iHeps, however, has hampered it uses in cell-based therapy. Since hepatic stem cells (HepSCs) possess self-renewal and bipotency with the capacity to differentiate into both hepatocytes and cholangiocytes, they have therapeutic potential for treating liver disease. Here, we investigated the therapeutic effects of induced HepSCs (iHepSCs) on a carbon tetrachloride (CCl4)-induced liver fibrosis model. We demonstrate that Oct4 and Hnf4a are sufficient to convert fibroblasts into expandable iHepSCs. Hepatocyte-like cells derived from iHepSCs (iHepSC-HEPs) exhibit the typical morphology of hepatocytes and hepatic functions, including glycogen storage, low-density lipoprotein (LDL) uptake, Indocyanine green (ICG) detoxification, drug metabolism, urea production, and albumin secretion. iHepSCs-derived cholangiocyte-like cells (iHepSC-CLCs) expressed cholangiocyte-specific markers and formed cysts and tubule-like structures with apical-basal polarity and secretory function in three-dimensional culture condition. Furthermore, iHepSCs showed anti-inflammatory and anti-fibrotic effects in CCl4-induced liver fibrosis. This study demonstrates that Oct4 and Hnf4α-induced HepSCs show typical hepatic and biliary functionality in vitro. It also presents the therapeutic effect of iHepSCs in liver fibrosis. Therefore, directly converting iHepSCs from somatic cells may facilitate the development of patient-specific cell-based therapy for chronic liver damage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbon Tetrachloride Poisoning* / genetics
  • Carbon Tetrachloride Poisoning* / metabolism
  • Carbon Tetrachloride Poisoning* / therapy
  • Hepatocyte Nuclear Factor 4* / genetics
  • Hepatocyte Nuclear Factor 4* / metabolism
  • Induced Pluripotent Stem Cells* / metabolism
  • Induced Pluripotent Stem Cells* / pathology
  • Induced Pluripotent Stem Cells* / transplantation
  • Liver Cirrhosis* / chemically induced
  • Liver Cirrhosis* / genetics
  • Liver Cirrhosis* / metabolism
  • Liver Cirrhosis* / therapy
  • Liver* / metabolism
  • Liver* / pathology
  • Lung Injury* / chemically induced
  • Lung Injury* / genetics
  • Lung Injury* / metabolism
  • Male
  • Mice
  • Octamer Transcription Factor-3* / genetics
  • Octamer Transcription Factor-3* / metabolism
  • Stem Cell Transplantation*

Substances

  • Hepatocyte Nuclear Factor 4
  • Hnf4a protein, mouse
  • Octamer Transcription Factor-3
  • Pou5f1 protein, mouse

Grants and funding

JBK received funding for this work by the Bio & Medical Technology Development Program of the National Research Foundation (NRF) funded by the Ministry of Science & ICT (Grant No. 2016M3A9B4919719 and Grant No. 2017M3A9C6033875) and Tech Incubator Program for Startup (TIPS) funded by the Ministry of SMEs and Startups (S2566811). The funders (www.nrf.re.kr) had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.