Differentiation and enrichment of hepatocyte-like cells from human embryonic stem cells in vitro and in vivo

YuYou Duan, Andreea Catana, Ying Meng, Naoki Yamamoto, Songqing He, Sanjeev Gupta, Sanjiv Sam Gambhir, Mark A Zern

Research output: Contribution to journalArticle

165 Citations (Scopus)

Abstract

Human embryonic stem cells (hESC) may provide a cell source for functional hepatocytes. The aim of this study is to establish a viable human hepatocyte-like cell line from hESC that can be used for cell-based therapies. The differentiated hESC were enriched by transducing with a lentivirus vector containing the green fluorescent protein (GFP) gene driven by the α1-antitrypsin promoter; the GFP gene is expressed in committed hepatocyte progenitors and hepatocytes. GFP+ hESC were purified by laser microdissection and pressure catapulting. In addition, differentiated hESC that were transduced with a lentivirus triple-fusion vector were transplanted into NODSCID mice, and the luciferase-induced bioluminescence in the livers was evaluated by a charge-coupled device camera. GFP+ hESC expressed a large series of liver-specific genes, and expression levels of these genes were significantly improved by purifying GFP+ hESC; our results demonstrated that purified differentiated hESC express nearly physiological levels of liver-specific genes and have liver-specific functions that are comparable to those of primary human hepatocytes. The differentiated hESC survived and engrafted in mouse livers, and human liver-specific mRNA and protein species were detected in the transplanted mouse liver and serum at 3 weeks after transplantation. This is the first time that human albumin generated by hESC-derived hepatocytes was detected in the serum of an animal model. This also represents the first successful transplantation of differentiated hESC in an animal liver and the first bioluminescence imaging of hESC in the liver. This study is an initial step in establishing a viable hepatocyte-like cell line from hESC.

Original languageEnglish (US)
Pages (from-to)3058-3068
Number of pages11
JournalStem Cells
Volume25
Issue number12
DOIs
StatePublished - Dec 2007

Fingerprint

Hepatocytes
Liver
Green Fluorescent Proteins
Lentivirus
In Vitro Techniques
Human Embryonic Stem Cells
Genes
Transplantation
Cell Line
Microdissection
Cell- and Tissue-Based Therapy
Luciferases
Serum
Albumins
Lasers
Animal Models
Gene Expression
Pressure
Equipment and Supplies
Messenger RNA

Keywords

  • Hepatic differentiation
  • Human embryonic stem cells
  • Laser microdissection and pressure catapulting Molecular imaging
  • Lentivirus vectors

ASJC Scopus subject areas

  • Cell Biology

Cite this

Differentiation and enrichment of hepatocyte-like cells from human embryonic stem cells in vitro and in vivo. / Duan, YuYou; Catana, Andreea; Meng, Ying; Yamamoto, Naoki; He, Songqing; Gupta, Sanjeev; Gambhir, Sanjiv Sam; Zern, Mark A.

In: Stem Cells, Vol. 25, No. 12, 12.2007, p. 3058-3068.

Research output: Contribution to journalArticle

Duan, Y, Catana, A, Meng, Y, Yamamoto, N, He, S, Gupta, S, Gambhir, SS & Zern, MA 2007, 'Differentiation and enrichment of hepatocyte-like cells from human embryonic stem cells in vitro and in vivo', Stem Cells, vol. 25, no. 12, pp. 3058-3068. https://doi.org/10.1634/stemcells.2007-0291
Duan, YuYou ; Catana, Andreea ; Meng, Ying ; Yamamoto, Naoki ; He, Songqing ; Gupta, Sanjeev ; Gambhir, Sanjiv Sam ; Zern, Mark A. / Differentiation and enrichment of hepatocyte-like cells from human embryonic stem cells in vitro and in vivo. In: Stem Cells. 2007 ; Vol. 25, No. 12. pp. 3058-3068.
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