Protocol for Directed Differentiation of Human Induced Pluripotent Stem Cells (iPSCs) to a Hepatic Lineage

Methods Mol Biol. 2017:1639:151-160. doi: 10.1007/978-1-4939-7163-3_15.

Abstract

Directed differentiation is a powerful cell culture technique where developmental pathways are applied to a pluripotent progenitor in order to generate specific terminally differentiated cell populations. Here, we describe a serum-free protocol using growth factors in defined concentrations to derive iPSC-hepatic cells starting from both feeder and feeder-free conditions. The generated iPSC-hepatic cells are developmentally similar to fetal stage hepatocytes, and when generated from patients with genetic mutations such as alpha-1 antitrypsin deficiency recapitulate pathologic changes associated with clinical disease, such as protein misfolding, intracellular retention of misfolded proteins, and elevated levels of ER stress.

Keywords: Definitive endoderm; Directed differentiation; Induced pluripotent stem cell (iPSC); iPSC-hepatic cell.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / methods*
  • Cell Differentiation*
  • Cell Lineage*
  • Cell Shape
  • Endoderm / cytology
  • Feeder Cells / cytology
  • Flow Cytometry
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Liver / cytology*
  • alpha 1-Antitrypsin Deficiency / metabolism