The analysis for proliferation and differentiation of hepatic stem cells

我们发现持续性损伤诱导bmc向功能性肝细胞的高效转分化。将四氯化碳诱导的肝硬化小鼠经高静脉注射1 × 10^5未处理的绿色荧光蛋白（GFP）阳性BMCs。在这些小鼠中，移植的gfp阳性BMCS在一天后有效地迁移到肝小叶的门静脉周围区域，并在4周内重新填充了受体肝脏的四分之一。相比之下，对照小鼠移植后未检测到gfp阳性bmc。肝脏完好无损。bmc通过未成熟的肝母细胞向功能成熟的肝细胞转化。血清白蛋白显著升高，以补偿BMC移植引起的慢性肝衰竭。这些结果表明，受体条件和微环境是BMC细胞治疗成功的关键因素。我们将此模型命名为GFP/CC14模型。下一步，我们开发了一种新的单克隆抗体，抗liv8抗体。在胚胎第11.5天，在缺乏造血系统的AML1敲除小鼠的肝脏中发现了liv8阳性细胞，但没有发现。成年小鼠BMC的32%为liv8阳性细胞，其中包括cd45阳性细胞。我们利用自动磁性细胞分选系统分离liv8阳性和liv8阴性细胞，然后将这些细胞移植到连续肝损伤模型中。一周后，接受liv8阳性细胞和liv8阴性细胞的肝脏中BMC的初始定植没有明显差异，但在4周时，liv8阴性细胞更有效地将BMC转分化为肝细胞。现在我们正试图识别抗liv8的抗原。这些分析将有助于开发所有有效的细胞疗法来修复受损的肝脏。另一方面，我们发现了这种新的蛋白质。Human homologue of maid （HHM）是一种参与肝干细胞发育和分化的螺旋-环-螺旋（HLH）转录调控蛋白。我们分析了HHM在肝癌发生中的潜在作用。我们通过原位杂交（原位杂交）和免疫组化分析在大鼠肝癌发生的氯缺乏l -氨基酸（CDAA）饮食模型和人类HCC样本中检测了HHM的表达。HHM在灶性和HCC中高表达。这些结果提示HHM可能是检测肝癌发生起始的一个有用的标记蛋白

Here we found that persistent injury induced efficient trans-differentiation of BMCs into functional hepatocytes. Liver cirrhosis mice induced by carbon tetrachloride were injected with 1x10^5 non-treated green fluorescent protein (GFP)-positive BMCs via tall vein. In these mice, transplanted GFP-positive BMCS efficiently migrated into the peri-portal area of liver lobules after one day and repopulated one-fourth of the recipient liver by 4 weeks. In contrast, no GFP-positive BMCs were detected following transplantation into control mice. with undamaged livers. BMCs trans-differentiated into functional mature hepatocytes via immature hepatoblasts. Serum albumin was significantly elevated to compensate for chronic liver failure by BMC transplantation. These results showed that the recipient condition and microenvironments are key factors for successful cell therapy using BMC. We named this model as a GFP/CC14 model. As the next step, we developed a new monoclonal antibody, anti-Liv8 usi … More ng mouse fetal liver antigen. On embryonic day 11.5, Liv8-positive cells were seen in the liver of wild-tape, but not AML1 knockout mice, which lack the hematopoietic system. Liv8-positive cells accounted for 32% of the BMC in adult mice and included CD45-positive cells. We separated Liv8-positive or Liv8-negative cells using Auto Magnetic Cell Sorting system, and then transplanted these cells to a continuous liver damaged model. After one week, there was no marked difference in the initial colonization of BMC in the liver between recipients of Liv8-positive and Liv8-negative cells, but at 4 weeks more efficient BMC trans-differentiation into hepatocytes was seen with Liv8-negative cells. Now we are trying to identify the antigen of anti-Liv8. These analysis will be useful to develop all efficient cell therapy to repair damaged liver. On the other hands, we identified that new protein. Human homologue of maid (HHM) is a helix-loop-helix (HLH) transcriptional regulatory protein that is involved in the hepatic stem cell development and differentlation. We analyzed the potential involvement of HHM in hepatocarcinogenesis. We assessed HHM expression in the chorine deficient L-amino acid defined (CDAA) diet model of rat hepatocarcinogenesis by in situ hybridization DISH), and human HCC samples by immunohistochemical analysis. High HHM expression was seen in foci and HCC. These results suggested that HHM may be a useful marker protein to detect initiation of hepatocarcinogenesis Less