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后肝脏明显增高。LEG数量在第7天最高；比PVL之前高15倍。PVL后，LEG主要出现在门静脉周围区域，在PVL后第7天，肝脏中菌落数量最多，说明PVL后LEG的分离效率较高。2)人肝门静脉栓塞后，LEG类AFP阳性细胞主要出现在门静脉周围区域。AFP阳性率与栓塞肺叶细胞凋亡的发生成正比，与未栓塞肺叶细胞动力学的发生成反比。3)采用不经门静脉灌注的分离方法可从肝脏中分离培养LEC，说明肝活检标本中可获得LEC。4)转染HNF-3a不能使LEC在体外分化为肝细胞。5)移植到NAR肝脏的LECs分化为产生白蛋白的肝细胞。6)脾脏移植的LECs经脾转位和门静脉分流后未分化为肝细胞。因此，肝栓塞后lec数量明显增加，门静脉栓塞后可以有效地从肝脏中分离出来。这些结果为临床PE后肝干细胞样细胞移植提供了可能性。

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.