Hepatic Differentiation of Embryonic Stem Cells by Murine Fetal Liver Mesenchymal Cells

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 946)

Abstract

Hepatocytes derived from embryonic stem cells (ESCs) are a potential cell source for regenerative medicine. However, it has been technically difficult to differentiate ESCs into mature hepatocytes because the definitive growth factors and molecular mechanisms governing hepatocyte differentiation have not yet been well defined. The CD45CD49f+/−Thy1+gp38+ mesenchymal cells that reside in murine fetal livers induce hepatic progenitor cells to differentiate into mature hepatocytes by direct cell–cell contact. Utilizing these cells, we employ a two-step procedure for hepatic maturation of ESCs: first, ESCs are differentiated into endodermal cells or hepatic progenitor cells, and second, ESC-derived endodermal cells are matured into functional hepatocytes by coculture with murine fetal liver mesenchymal cells. The ESC-derived hepatocyte-like cells possess hepatic functions, including ammonia removal activity, albumin secretion ability, glycogen synthesis and storage, and cytochrome P450 enzymatic activity.

Key words

Embryonic stem cell Fetal liver Hepatocyte Hepatic progenitor cell Mesenchymal cell Thy1 gp38 

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Takamichi Ishii
    • 1
  • Kentaro Yasuchika
    • 1
  • Iwao Ikai
    • 1
    • 2
  1. 1.Department of Surgery, Graduate School of MedicineKyoto UniversityKyotoJapan
  2. 2.Department of Surgery, Kyoto Medical CenterNational Hospital OrganizationKyotoJapan

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