Study of analysis of bile stasis using the extracorporeal hepatic tissue model and of the effect of cell transplantation on hepatic failure

本研究计划由三部分组成，即第一部分“体外肝组织结构的建立”，第二部分“体外条件下胆汁淤积的研究”，第三部分“细胞移植治疗肝功能衰竭”。第一个是最重要的研究，延伸到再生医学和人工肝。小肝细胞培养诱导出类似肝脏、组织的类器官结构。此外，小肝细胞即使在长期冷冻保存后仍能保持生长能力和肝脏分化功能。这一结果意味着建立小鼠和人肝细胞的分离方法将是重要的，该方法的应用可能对建立肝细胞库和人工肝的发展有所帮助。第二项研究是通过细胞因子治疗的对联中胆小管收缩的数量来进行的。The ra…经肿瘤坏死因子-α处理的肝细胞偶联体细胞间隙内可见双侧小管内的液滴有更明显的内源性运动。肿瘤坏死因子-α和IL-6可减少胆小管收缩。此外，在大鼠小肝细胞和非实质细胞重建的肝脏器官结构中形成的胆小管的运动也有类似的结果，这些肝细胞的利用是指细胞移植到肝脏中。在某些情况下可以观察到注射的肝细胞成功增殖，但实验中的重复性结果尚未确定。总之，小肝细胞的培养提供了类肝细胞的结构，促进了肝细胞的分化，并保持了组织特异性计划的典型特征。这可能会对肝功能衰竭患者的护理产生重大的治疗影响。较少

This research program was composed of three components, namely the first is"the establishment extracorporeal hepatic tissue structure", the second is"the study of bile stasis under in vitro condition" and the third is "the treatment of hepatic failure by cell transplantation". The first one is most important research extending to the regenerative medicine and to the tartificial liver. The culture of small hepatocyte invited the organoid structure resembling with liver, tissue. Moreover, small hepatocytes maintain growth ability and hepatic differentiated functions even after long-term cryopreservation. This result means that the establishment of isolation method for mouse and human hepatocytes will be important, and the application of the method shown in this study may be useful for the establishment of a hepatocyte bank and development of an artificial liver. The second research was performed by the number of bile canalicular contractions in the couplets treated with cytokines. The ra … More pid movement of a droplet from bilecanaliculus was observed at the intercellular space of the hepatocyte couplet treated with TNF-α. And TNF-αand IL-6 reduce bile canaliculus contractions. Moreover, the similar result was observed in the movement of bile canaliculus formed in hepatic organoid structure reconstructed by rat small hepatocytes and nonparenchymal cells.The utilization of those hepatic cells referred to cell transplantation into a liver. The successful proliferation of the injected hepatocyte observed in some cases, but reproducible results in experiments have not been established. Further efforts would be required for the establishment of hepatocytic stem cell transplantation in liver tissue.In summary, we demonstrated that, the culture of small hepatocyte provides a hepatoid structure, improves hepatocyte differentiation and maintains a tissue-specific plans typical characteristics. This might have a significant therapeutic impact on the care of the patients with liver failure. Less