J Korean Soc Transplant.  2007 Dec;21(2):216-222.

In vitro Differentiation of Human Embryonic Stem Cells into Definitive Endodermal Cells

• Lim M
• Choi D
• Kim SJ
• Cheong HJ
• Won JH

Abstract

PURPOSE: Whole liver transplantation has limitation including donor shortage and fatal surgical complications. Hepatocyte transplantation, which is simpler and less expensive than whole liver transplantation, allows the use of living related donors, permits the use of a single donor organ for multiple recipients. However, hepatocytes have limitation in proliferation and lose their property during culture period. To over come these problems, here we performed differentiation of human embryonic stem cells (hESCs) into definitive endoderm in order to differentiate into hepatocytes efficiently.
METHODS
Undifferentiated hESCs were maintained on mouse embryo fibroblast feeder (MEF) layer for 5~7 days. For endoderm differentiation, we used modified Kevin A D'Amour's method that added 100 ng/mL Activin A for 5 days. After differentiation, differentiated endodermal cells were collected and RT-PCR and immunostain analysis were performed.
RESULTS
After 5 days of differentiation period, hES cells showed endoderm committed-cells and increased expression of endoderm-specific marker genes (Sox17 and Foxa2). Also differentiated endoderm cells were stained with Sox17 and Foxa2 whereas undifferentiated hES cells were not stained with Sox17, Foxa2.
CONCLUSION
In vitro differentiotion from hES cells to definitive endoderm was done repetitively by our methods. Further well defined protocol for differentiation of definitive endoderm to hepatocytes should be made.

Keyword

Human embryonic stem cells (hESCs); Definitive endoderm

MeSH Terms

Activins
Animals
Embryonic Stem Cells*
Embryonic Structures
Endoderm*
Fibroblasts
Hepatocytes
Humans*
Liver Transplantation
Mice
Tissue Donors
Activins