Treatment of surgically induced acute liver failure by transplantation of HNF4-overexpressing embryonic stem cells.
OBJECTIVE
Tissue-specific stem cells from differentiating embryonic stem (ES) cells are both pluripotent and genetically flexible. Recent observations indicate that ES cells can differentiate into hepatocytes. Therefore, cell-based therapy can potentially be a therapeutic alternative to liver transplantation. In this study the treatment of acute liver failure in rats by transplantation of hepatocyte nuclear factor 4 (HNF4)-overexpressing ES cells was investigated.


METHODS
The HNF4 was transfected into ES cells and ES cell clones overexpressing HNF4 were selected. The levels of markers of hepatocyte differentiation, including albumin, transthyretin, glucose-6-phosphates (G-6-P) and SAPK/ERK kinase-1 (SEK1) mRNA, were tested in spontaneously differentiated HNF4-overexpressing ES cells by reverse transcription-polymerase chain reaction (RT-PCR). The ultrastructure of the spontaneously differentiated HNF4-overexpressing ES cells was examined by electron microscopy. To induce acute liver failure, Sprague-Dawley rats were subjected to 90% hepatectomy and given 5% oral dextrose. The rats were divided into three groups. The rats in the treatment group (n = 12) received intraliver injection of 2 x 10(7) undifferentiated HNF4-overexpressing ES cells from the same clone, the rats in control group 1 (n = 12) received 2 x 10(7) undifferentiated ES cells, and the rats in control group 2 (n = 12) received the same volume of media without any cells.


RESULTS
All rats in control group 1 and control group 2 died within 72 h, while 33% of rats that received undifferentiated HNF4-overexpressing ES cells transplantation survived more than 1 month. Spontaneously differentiated HNF4-overexpressing ES cells only expressed transthyretin mRNA. The cells were rich in mitochondrion and catalase-containing peroxisomes in ultrastructure.


CONCLUSIONS
Transplantation of ES cells could be a potential treatment in supporting life during acute liver insufficiency and could be a bridge to orthotopic liver transplantation.
Kuai
Xiao Ling
XL
Shanghai Institute of Digestive Diseases, Renji Hospital, Shanghai Second Medical University, 145 Shandong Zhong Road, Shanghai 200001, China.
Cong
Xiao Qian
XQ
Du
Zhong Wei
ZW
Bian
Yu Hai
YH
Xiao
Shu Dong
SD
D000818:Animals; D015415:Biomarkers / Q000097:blood; D002454:Cell Differentiation; D004195:Disease Models, Animal; D006498:Hepatectomy / Q000009:adverse effects*; D051557:Hepatocyte Nuclear Factor 4 / Q000096:biosynthesis* / Q000627:therapeutic use* / Q000648:ultrastructure; D022781:Hepatocytes / Q000378:metabolism / Q000648:ultrastructure; D017114:Liver Failure, Acute / Q000209:etiology* / Q000401:mortality / Q000473:pathology / Q000628:therapy*; D008297:Male; D008930:Mitochondria, Liver / Q000378:metabolism / Q000648:ultrastructure; D020675:Peroxisomes / Q000378:metabolism / Q000648:ultrastructure; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000378:metabolism / Q000648:ultrastructure; D015854:Up-Regulation
D015415:Biomarkers; D051557:Hepatocyte Nuclear Factor 4; D012333:RNA, Messenger