Embryonic or Induced Pluripotent Stem Cell-Derived Hepatocellular Systems for HCV Culture

Methods Mol Biol. 2019:1911:121-135. doi: 10.1007/978-1-4939-8976-8_8.

Abstract

Human-induced pluripotent stem cell-derived hepatocyte-like cells (iHeps) constitute a powerful tool for modeling hepatotropic pathogen infections in cell culture. Meanwhile, CRISPR-Cas9 technology enables precise editing of stem cell genomes to generate patient-specific disease models and thus development of personalized experimental systems. Here we present a detailed stepwise protocol for the differentiation of stem cells to hepatocyte-like cells for HCV studies in cell culture. We also outline the use of an inducible iCRISPR platform for the rapid and efficient modification of host factors of interest to better understand their function during HCV infection.

Keywords: Hepatocyte-like cells; Human embryonic/induced pluripotent stem cells; Precision medicine; iCRISPR.

MeSH terms

  • CRISPR-Cas Systems
  • Cell Culture Techniques / methods*
  • Cell Differentiation*
  • Cell Line
  • Gene Editing / methods
  • Hepacivirus / physiology
  • Hepatitis C / genetics
  • Hepatocytes / cytology*
  • Hepatocytes / metabolism
  • Host-Pathogen Interactions
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism