[The expression of human specific proteins in liver tissue of chimeric goats engrafted with human hematopoitic stem cells].
OBJECTIVE
To investigate the expression of human specific proteins in liver tissue obtained from goats engrafted with human hematopoietic stem cells (hHSC).


METHODS
hHSCs derived from cord blood were transplanted into fetal goats for production of chimerism. Liver specimens were obtained from four 10-month-old chimeric goats and examined for their reactivity with monoclonal antibodies against human antigens: proliferating cell nuclear antigen (PCNA) and hepatocyte specific antigen (HSA). The presence of human HSA positive cells in the goat liver tissue was determined by fluorescence assisted cell sorting (FACS). The expression of human hepatocyte nuclear factor (hHNF-3beta) and human serum albumin (hALB) mRNAs was determined by RT-PCR. Meanwhile, FISH experiment was also performed to detect the human cells in the chimeric livers with the probe of human p17H8.


RESULTS
Successful engraftment was confirmed by the detection of human blood cell chimerism in the circulation of the transplanted goats. Human PCNA and HSA were found in liver specimens of transplanted goats but not of normals. FACS analysis showed the presence of human HSA positive cells in the liver of chimeric goats. RT-PCR results demonstrated that hHNF-3beta and hALB mRNAs were specifically expressed in liver tissues of chimeric goats. FISH experiment showed two positive hybridized signals in some liver cells of the chimeric goats, indicating that human liver-like cells were present there.


CONCLUSIONS
Goats engrafted with hHSC are capable to produce chimeric livers. Growth and propagation of human cells in the liver tissue of such transplanted goats were possible. Particularly, the human liver-like cells in chimeric goat livers had transcriptional activity specific to human hepatocytes.
Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai 200040, China.
Shuzhen
S
Huang
Hinfai
H
Yam
Chipui
C
Pang
Meijue
M
Chen
Zhijuan
Z
Gong
Fanyi
F
Zeng
Shunchiu
S
Ling
Yitao
Y
Zeng
D000818:Animals; D004268:DNA-Binding Proteins / Q000235:genetics*; D006041:Goats; D051542:Hepatocyte Nuclear Factor 3-beta; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D008099:Liver / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D012709:Serum Albumin / Q000235:genetics*; D033581:Stem Cell Transplantation; D014157:Transcription Factors; D018183:Transplantation Chimera / Q000378:metabolism*
D004268:DNA-Binding Proteins; C496502:FOXA2 protein, human; D009687:Nuclear Proteins; D012333:RNA, Messenger; D012709:Serum Albumin; D014157:Transcription Factors; D051542:Hepatocyte Nuclear Factor 3-beta