[Study of embryonic stem cells induced to express hepatic cell functions in vitro in a pathologic niche containing cholestatic serum].
OBJECTIVE
To study the role of a pathologic niche inducing mouse embryonic stem cells (ESC) to express hepatic cell functions in vitro.


METHODS
Embryoid bodies were developed from 5 to 7 day hanging-drop culture of mouse ESC, and their dissociated cells were planted in three differential systems: nothing added; with 20 ng/ml hepatocyte growth factor (HGF); and 5% rat cholestatic serum plus 20 ng/ml HGF added. Their differentiation was observed with inverted microscopes daily, and their hepatic functions were analyzed against their synthesis of glycogen, triglycerides, albumin, and urea nitrogen, and by their staining of indocyanine green (ICG) and fluorescein diacetate (FDA).


RESULTS
ESC spontaneous differentiation was hardly being controlled to form three germ layers. HGF prompted the ESC to develop further into visceral endoderm and mesoderm (myocardium), but both of them only expressed a low level of hepatocyte-specific metabolic functions. With cholestatic serum added into the HGF-induced system, differentiated cells grew into similar angular cells, and had a higher level synthesis of glycogen, triglycerides, albumin and urea nitrogen with positive ICG and FDA staining.


CONCLUSIONS
Spontaneous or HGF-induced ESC differentiation has only limited hepatic functions expressed. A pathologic niche in vitro induces ESC to develop into hepatic lineages, with a higher level of hepatic metabolic functions.
Fang
Tian-Ling
TL
Hepatobillary Surgery, Center of Medical Research, the Second Affiliated Hospital, Sun Yet-sen University, Guangzhou 510120, China.
Min
Jun
J
Deng
Xiao-Geng
XG
Qian
Shi-Kun
SK
Chu
Zhong-Hua
ZH
Chen
Ya-Jin
YJ
Shao
Jing
J
Wei
Jing
J
Chen
Ji-Sheng
JS
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D002779:Cholestasis / Q000097:blood*; D003470:Culture Media / Q000494:pharmacology; D004622:Embryo, Mammalian; D022781:Hepatocytes / Q000166:cytology*; D051379:Mice; D044967:Serum; D013234:Stem Cells / Q000166:cytology*
D003470:Culture Media