چكيده لاتين :
Human embryonic stem cells (hESC), which are derived from the inner cell mass of the
blastocysts, have been considered to be pluripotent cells. In this study we examine the differentiating potential
of hESC into hepatocytes by characterization of the expression of endoderm and liver-specific genes.
Methods: hESC were cultivated in suspension to form aggregates, the embryoid bodies. They were allowed to
outgrowth on the plated culture with the stepwise addition of growth factors such as acidic fibroblast growth
factor (aFGF), hepatocyte growth factor and oncostatin M into the culture medium. The expressions of
endodermal and liver specific genes such as hepatocyte nuclear factor 3p, alpha-fetoprotein (AFP), albumin
(ALB), cytokeratin 8 (CK-8), CK-18, transthyretin, glucose 6-phosphatase and tyrosine aminotransferase were
analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The expressions of ALB and CK-18
in the cytoplasm were analyzed by Immunohistochemistry. Results: The immunoblotting and
chemiluminescence of the conditioned media indicated the secretion of ALB and AFP. RT-PCR analysis
revealed that hepatic gene expression related to early and late-stage liver development were enhanced through
in vitro differentiation of hESC. Conclusion: Our results showed the expression of endoderm and hepatic
specific genes after in vitro differentiation of hESC into hepatocyte-like cells through addition of various
growth factors in three dimensional culture systems (collagen type I). hESC could be a new potential source of
hepatocyte for transplantation in patients with liver failure. Iran. Biomed. J. 10 (3): 117-124, 2006
