Differentiation of hepatic stem cells into hepatocytes by biotechnology and those transplantation to the liver-like spleen with diversion of portal blood flow

1)PVL后肝组织明显增多。术后第7天小腿数最多，是术前的15倍。PVL后小腿主要出现在门静脉周围区域，PVL后7d肝内集落形成最多，提示PVL后分离小腿的效率较高。AFP阳性率与肝动脉栓塞区的细胞凋亡率成正比，与非栓塞区的细胞动力学发生率成反比。3)无门脉灌流的分离方法可从肝脏分离培养出LEC，提示肝活检组织可获得LEC。4)HNF-3a基因导入LEC不能使LEC在体外分化为肝细胞。5)移植到NAR肝脏的LEC在体外分化为产生白蛋白的肝细胞。6)移植到脾内的LEC在脾移位和门静脉血流改道后不能分化为肝细胞。PE后LEC数量明显增多，门静脉栓塞术后LEC可有效地从肝脏分离出来。这些结果为临床进行PE后肝干细胞样细胞移植提供了可能。

1) There was a marked increase in the liver after PVL. The number of LEG was highest on day 7; 15 times higher than that before PVL. LEG was predominantly appeared in the periportal area after PVL, The number of colonies was highest from the liver on day 7 after PVL, which implicated efficiency in isolation of LEG after PVL.2) LEG like AFP positive cells appeared predominantly in the periportal area of human liver after portal embolization. AFP positive rate was proportional to occurrence of apoptosis in the embolized lobe and inversely proportional to occurrence of cell kinetics in the non-embolized lobe.3) LEC could be isolated and cultured from the liver by the isolation procedure without portal perfusion, which implicates that LEC could be obtained from the liver biopsy specimen.4) The transfection of HNF-3a into LEC could not make LEC differentiate into hepatocytes in vitro.5) LECs transplanted to the liver of NAR differentiated into hepatocytes that produced albumin.6) LECs transplanted to the spleen did not differentiate into hepatocytes after the splenic transposition and portal flow diversion.Consequently, LECs were markedly increased in number after PE and could efficiently be isolated from the liver after portal embolization. These results open up a possibility in cellular transplantation of hepatic stem-like cells from the liver after PE in clinical settings.