| Objective: To study the effect of liver cell xenograft in nude mice small forsize syndrome model which80%liver tissue is resected. The liver function,mortality rate, survival time and incidence of small liver syndrome areinvestigated.Methods: Nude mice (5weeks, weight22.71±1.11g) are forbidden food andwater for6hours before surgery.1%pentobarbital (0.01ml/g) is given to nudemice by intraperitoneal injection. The liver is exposed by abdominal midlineincision.80%liver tissue is resected for small for size liver syndrome modelestablishment. Then, the nude mice suffered operations are randomly dividedinto two groups. One is pig liver cell transplantation group; another is controlgroup,10cases in each group. Small pig is selected for liver cell donor. Pigliver lobe is cut and made to liver cell suspension. Pig liver cell suspension isinjected into nude mice of pig liver cell transplantation group by intraperitoneal(1ml liver cell suspension in each nude mice, the liver cell number is to be1x106).1ml of saline is injected into abdominal cavity of nude mice in controlgroup. The survival time and survival condition of each nude mice are observed.0.1ml blood is drawn by heart puncture for serum ALT、AST values test at1hbefore transplantation, on1d,4d,7d,14d, and28d after transplantationrespectively. Liver biopsy and pig liver cell xenograft biopsy in nude mice aredone at1h before transplantation, on2d,7d,28d and56d after transplantation respectively. The liver tissue slices are stained by HE staining to observe thepathological change of hepatocytes and xenograft.Results①Porcine hepatocytes were harvested by collagenase perfusionmethod which survival rate was more than90%confirmed by trypan bluedyeing. On56d after transplant, scattered porcine hepatocyte mass can be seenin the abdominal cavity of nude mice in transplant group which confirmed bybiopsy.②On56d after transplantation, the survival rate of nude mice in thecontrol group was20%, while the survival rate of nude mice in the transplantgroup was80%.③ALT, AST of nude mice in transplant group hadsignificant rise in value before transplantation, respectively ALT525.22±28.21U/L, AST428.32±35.45U/L, which to small for size liver syndrome diagnosticcriteria. Liver function significantly decreased on1d postoperative. ALT andAST of nude mice in control group were respectively523.13±21.35U/L,426.54±32.65U/L before transplantation. It began to gradually decline on4dafter transplantation. ALT, AST of nude mice in transplant group weresignificant less than that in control group on1d after transplantation (P <0.05).④The liver tissue in each nude mouse was partly missing before transplantation;the color of liver was a bit dark. Swelling and steatosis liver tissue wasobserved in control group on2d after transplantation, abdominal small turbidascites. The liver tissue appeared red colored, texture soft in transplantationgroup, blood stasis become lighter, swelling and fat samples without ascites.The liver tissue in control group became a ruddy, slightly soft texture on7d after transplantation, ascites was still visible. Liver tissue regeneration wasobserved in transplant group on7d after transplantation, the regenerate livertissue appeared texture soft, color is ruddy. Liver tissue regeneration could beseen in both transplant group and control group on28d after transplantation.The liver tissues in them were soft texture, color and luster is ruddy. The liverof nude mice in both two groups appeared normal on56d after transplantation.The liver weight of nude mice increased.⑤On2d and7d after porcinehepatocytes transplantation, in the liver of nude mice in control group, centralvein was severe congestion, hepatocyte ballooning became apparent,hepatocyte was swelling, large hepatocytes appeared necrosis, hepatic cablestructure disappeared with a large amount of inflammatory cell infiltration.However, the liver tissue of nude mice in transplant group only appeared asmall amount of cavitations and became a little fatty, necrosis of hepatocyteswere less. There was still basic normal hepatic and portal area structure.Normal liver cable structure and lobular architecture could be seen in the liverof nude mice in transplant group on56d after transplantation. The hepaticlobule structure in the liver of nude mice in control group returned to normal on56d after transplantation, but liver cable structure was still disorder.Conclusions: porcine hepatocyte xenograft in the abdominal cavity of nudemice can well survive still for56days. Pig hepatocyte xenograft can improvethe liver function of nude mice after80%liver resection; it can reduce themortality of nude mice and the incidence of liver failure. It can build a bridge between small-for-size syndrome after most of liver resection and liverregeneration and recovery in nude mice. |
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