Diminution of toxic copper accumulation in toxic milk mice modeling Wilson disease by embryonic hepatocyte intrasplenic transplantation.
OBJECTIVE
To observe the therapeutic effect of intrasplenic transplantation with embryonic hepatocytes on amelioration of hereditary copper accumulation in toxic milk (TX) mouse modeling Wilson disease.


METHODS
Donor hepatocytes were harvested from 14-d fetal liver of a pregnant homogeneous DL mouse. These cells were successively cultured, labeled with fluorescein dye Hoechst 33342 for 24 h, and sequentially infused into the spleen parenchyma of the recipient TX mice. No host immunosuppression measures were taken. Two and four weeks after transplantation, the recipients were killed for routine histologic investigation and immunohistochemistry study up to 4 wk after transplantation. The serum copper and ceruloplasmin concentrations of the recipient mice were determined by graphite furnace atomic absorption spectroscopy.


RESULTS
In the following 2nd and 4th wk after transplantation, the donor hepatocytes could be visualized in the livers of 47.3% recipients. The serum ceruloplasmin and copper concentrations increased by 1.6-fold after 2 wk and 2.0-fold times after 4 wk respectively, which ultimately rose from about 30% of the normal level to nearly 60% (P<0.01). The hepatic copper concentration decreased 7.2%, 4 wk after transplantation. Pathologic examination showed that there were many actively proliferative hepatocyte precursor cells with specific embryonic hepatocyte marker AFP migrated into hepatic sinusoids of the recipients. A large number of cells carrying hepatocytes marker and albumin were observed in the recipient spleen tissues.


CONCLUSIONS
Embryonic hepatocytes are capable of differentiating into mature hepatocytes in vivo. After transplantation, the hereditary abnormalities of copper metabolism in TX mice could be corrected partially by intrasplenic transplantation of homogeneous embryonic hepatocytes.
Shi
Zhu
Z
Department of Neurology, Nanfang Hospital, 1st Military Medical University, Guangzhou 510515, Guangdong Province, China. junyishy@sohu.com
Liang
Xiu-Ling
XL
Lu
Bing-Xun
BX
Pan
Su-Yue
SY
Chen
Xi
X
Tang
Qi-Qiang
QQ
Wang
Ying
Y
Huang
Fan
F
D000818:Animals; D003300:Copper / Q000097:blood / Q000633:toxicity*; D004195:Disease Models, Animal; D005260:Female; D016332:Fetal Tissue Transplantation; D022781:Hepatocytes / Q000637:transplantation*; D006527:Hepatolenticular Degeneration / Q000139:chemically induced / Q000473:pathology / Q000628:therapy*; D008297:Male; D051379:Mice; D008892:Milk; D011247:Pregnancy; D013154:Spleen
D003300:Copper