The differentiation of hepatocyte-like cells from monkey embryonic stem cells

Xiaocui Ma, YuYou Duan, Christine J. Jung, Jian Wu, Catherine A. VandeVoort, Mark A Zern

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Embryonic stem cells (ESC) hold great potential for the treatment of liver diseases. Here, we report the differentiation of rhesus macaque ESC along a hepatocyte lineage. The undifferentiated monkey ESC line, ORMES-6, was cultured in an optimal culture condition in an effort to differentiate them into hepatocyte-like cells in vitro. The functional efficacy of the differentiated hepatic cells was evaluated using RT-PCR for the expression of hepatocyte specific genes, and Western blot analysis and immunocytochemistry for hepatic proteins such as α-fetoprotein (AFP), albumin and α1-antitrypsin (α1-AT). Functional assays were performed using the periodic acid schiff (PAS) reaction and ELISA. The final yield of ESC-derived hepatocyte-like cells was measured by flow cytometry for cells that were transduced with a liver-specific lentivirus vector containing the α1-AT promoter driving the expression of green fluorescence protein (GFP). The treatment of monkey ESC with an optimal culture condition yielded hepatocyte-like cells that expressed albumin, α1-AT, AFP, hepatocyte nuclear factor 3β, glucose-6-phophatase, and cytochrome P450 genes and proteins as determined by RT-PCR and Western blot analysis. Immunofluorescent staining showed the cells positive for albumin, AFP, and α1-AT. PAS staining demonstrated that the differentiated cells showed hepatocyte functional activity. Albumin could be detected in the medium after 20 days of differentiation. Flow cytometry data showed that 6.5 ± 1.0% of the total differentiated cells were positive for GFP. These results suggest that by using a specific, empirically determined, culture condition, we were able to direct monkey ESC toward a hepatocyte lineage.

Original languageEnglish (US)
Pages (from-to)485-493
Number of pages9
JournalCloning and Stem Cells
Volume10
Issue number4
DOIs
StatePublished - Dec 1 2008

ASJC Scopus subject areas

  • Biotechnology
  • Developmental Biology

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