Development of immortalized hepatocyte-like cells from hMSCs

Methods Mol Biol. 2012:826:73-87. doi: 10.1007/978-1-61779-468-1_7.

Abstract

Clones of hepatocyte-like cells were reproducibly generated from human mesenchymal stem cells immortalized with a combined transduction of both Bmi-1 and TERT genes. These hepatocyte-like cells contained selective markers and several functional properties of hepatocytes, yet still carried proliferative potential. These cells had cuboidal morphology and arranged themselves as cord-like structure in culture. The cloned cells deposited glycogen and actively synthesized albumin. The basal expressions of CYP450 isozymes was observed, albeit only 10-20% that of primary hepatocytes. These expressions were promptly increased upon the addition of rifampicin, a known enzyme inducer. These hepatocyte-like cells may serve as a close alternative to the use of primary hepatocytes for in vitro studies.

MeSH terms

  • Cell Culture Techniques / methods*
  • Cell Differentiation / physiology*
  • Cell Proliferation
  • Cell Separation / methods
  • Cell Shape / physiology
  • Cytochrome P-450 Enzyme System / metabolism
  • Enzyme Induction / physiology
  • Fluorescent Antibody Technique
  • Hepatocytes / cytology*
  • Humans
  • Isoenzymes / metabolism
  • Mesenchymal Stem Cells / cytology*
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Polycomb Repressive Complex 1
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism
  • Real-Time Polymerase Chain Reaction
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • Telomerase / genetics
  • Telomerase / metabolism
  • Transduction, Genetic

Substances

  • BMI1 protein, human
  • Isoenzymes
  • Nuclear Proteins
  • Proto-Oncogene Proteins
  • Repressor Proteins
  • Cytochrome P-450 Enzyme System
  • Polycomb Repressive Complex 1
  • TERT protein, human
  • Telomerase