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a Genome Institute of Singapore and the
b Departments of Pathology and
c Obstetrics and Gynaecology of the Faculty of Medicine, The National University of Singapore, Singapore;
d Transgenic/Gene Targeting Facility, Herbert Irving Comprehensive Cancer Center, Columbia University, New York, New York, USA
Key Words. Alpha-fetoprotein • Green fluorescent protein • Embryonic stem cells • Embryoid bodies • Hepatocytes
Sai-Kiang Lim, Ph.D., Genome Institute of Singapore, The National University of Singapore, 1 Science Park Road, The Capricorn #05-01, Singapore Science Park II, Singapore 117528, Singapore. Telephone: 65-6874-5247; Fax: 65-6773-5461; e-mail: gislimsk{at}nus.edu.sg
A major problem in gene therapy and tissue replacement is accessibility of tissue-specific stem cells. One solution is to isolate tissue-specific stem cells from differentiating embryonic stem (ES) cells. Here, we show that liver progenitor cells can be purified from differentiated ES cells using alpha-fetoprotein (AFP) as a marker. By knocking the green fluorescent protein (GFP) gene into the AFP locus of ES cells and differentiating the modified ES cells in vitro, a subpopulation of GFP+ and AFP-expressing cells was generated. When transplanted into partially hepatectomized lacZ-positive ROSA26 mice, GFP+ cells engrafted and differentiated into lacZ-negative and albumin-positive hepatocytes. Differentiation into hepatocytes also occurred after transplantation of GFP+ cells in apolipoprotein-E- (ApoE) or haptoglobin-deficient mice as demonstrated by the presence of ApoE-positive hepatocytes and ApoE mRNA in the liver of ApoE-deficient mice or by haptoglobin in the serum and haptoglobin mRNA in the liver of haptoglobin-deficient mice. This study describes the first isolation of ES-cell-derived liver progenitor cells that are viable mediators of liver-specific functions in vivo.
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