Induced maturation of hepatic progenitor cells in vitro

Braz J Med Biol Res. 2013 Jul;46(7):559-66. doi: 10.1590/1414-431X20132455. Epub 2013 Jul 16.


Hepatic progenitor cells (HPCs) are a potential cell source for liver cell transplantation but do not function like mature liver cells. We sought an effective and reliable method to induce HPC maturation. An immortalized HP14.5 albumin promoter-driven Gaussian luciferase (ALB-GLuc) cell line was established from HPCs isolated from fetal mouse liver of post coitus day 14.5 mice to investigate the effect of induction factors on ALB promoter. HP14.5 parental cells were cultured in DMEM with different combinations of 2% horse serum (HS), 0.1 µM dexamethasone (DEX), 10 ng/mL hepatic growth factor (HGF), and/or 20 ng/mL fibroblast growth factor 4 (FGF4). Trypan blue and crystal violet staining were used to assess cell proliferation with different induction conditions. Expression of hepatic markers was measured by semi-quantitative RT-PCR, Western blot, and immunofluorescence. Glycogen storage and metabolism were detected by periodic acid-Schiff and indocyanine green (ICG) staining. GLuc activity indicated ALB expression. The combination of 2% HS+0.1 µM Dex+10 ng/mL HGF+20 ng/mL FGF4 induced the highest ALB-GLuc activity. Cell proliferation decreased in 2% HS but increased by adding FGF4. Upon induction, and consistent with hepatocyte development, DLK, AFP, and CK19 expression decreased, while ALB, CK18, and UGT1A expression increased. The maturity markers tyrosine aminotransferase and apolipoprotein B were detected at days 3 and 6 post-induction, respectively. ICG uptake and glycogen synthesis were detectable at day 6 and increased over time. Therefore, we demonstrated that HPCs were induced to differentiate into functional mature hepatocytes in vitro, suggesting that factor-treated HPCs may be further explored as a means of liver cell transplantation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Differentiation / analysis
  • Apolipoprotein B-100
  • Apolipoproteins B / isolation & purification
  • Cell Differentiation / drug effects*
  • Cell Proliferation
  • Dexamethasone / administration & dosage
  • Embryo, Mammalian / drug effects*
  • Fibroblast Growth Factors / administration & dosage
  • Gentian Violet
  • Glycogen / metabolism
  • Hepatocyte Growth Factor / administration & dosage
  • Hepatocytes / cytology*
  • Indocyanine Green / pharmacokinetics
  • Liver / cytology*
  • Mice
  • Primary Cell Culture / methods
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stem Cells / cytology
  • Stem Cells / drug effects*
  • Trypan Blue
  • Tyrosine Transaminase / isolation & purification


  • Antigens, Differentiation
  • Apob protein, mouse
  • Apolipoprotein B-100
  • Apolipoproteins B
  • Fibroblast Growth Factors
  • Hepatocyte Growth Factor
  • Dexamethasone
  • Glycogen
  • Tyrosine Transaminase
  • Trypan Blue
  • Indocyanine Green
  • Gentian Violet