DEVELOPMENT OF TRANSPLANTATION REPLACEMENT TREATMENT BY IN UTERO TRANSPLANTATION OF ALLOGENEIC AND XENOGENEIC CELLS INTO FETAL LIVER

随着治疗方法的改进和临床后果的显著改善，器官移植已被广泛接受为治疗终末期患者的常规选择之一。相比之下，细胞移植和组织工程构建尚未被临床医生接受，尽管这一概念提出得很早。限制临床应用的主要因素之一是移植物在被宿主同化之前被排除在外。不仅是免疫介导的宿主反应，而且非特异性消除机制也被认为在移植物排斥反应中起着重要作用。尽管肝细胞损伤后有很好的再生潜力，但大多数终末期疾病，如重型肝炎或肝硬变，肝脏很难恢复。这些患者除了肝移植外无法治愈。在这里，同种异体肝细胞移植和人源化家畜肝脏t…在供体短缺的严峻形势下，更多的再移植作为一种新的治疗方法和新的供体来源受到了极大的关注。为了跨越异种细胞屏障，使人类细胞能够在猪器官内很好地固定和增殖，即使在免疫缺陷的猪中，也有必要了解宿主来源的非特异性消除系统的机制，并提高移植细胞的增殖能力。因此，本研究的目的是探讨异种细胞移植如何在猪肝内固定和增殖。基于腺病毒的p21转基因是一种已知的阻止细胞周期的分子，被转移到CB17SCID小鼠的肝脏中。在诱导p21转基因过表达的小鼠中完成部分肝切除。我们的数据表明，p21基因转基因小鼠部分切除后肝再生的潜力下降，通过评估肝脏重量和DNA合成的变化。THLE5b细胞可作为人肝实质细胞来源，体外增殖良好，但在CB17SCID小鼠体内仅能保持其正常形态特征。然而，在部分切除的p21转基因小鼠的肝脏中，THLE5b细胞开始增殖。我们的研究表明，p21基因转移对肝部分切除后的肝再生有负面影响，它可以为异种细胞的增殖提供一个合适的环境。较少

With the development of improved therapies and the significant advances in clinical consequence, organ transplantation has been widely accepted as one of the routine options for treatment of end-stage patients. In contrast, cell transplantation and tissue-engineered constructs have not been accepted by clinicians yet, even though the concept was proposed far early. One of the primary factors restricting the clinical application is the fact that grafts are excluded before they are assimilated in the host. Not only the immune-mediated host reaction, but non-specific elimination mechanism has been considered to play an important role contributing to the graft exclusion. Although it has been imaged that hepatocyte is of well potential to regenerate after injury, liver could hardly recover from most of end-stage disorders like severe hepatitis or cirrhosis. Those patients can be not cured except liver transplantation. Here, allogeneic hepatocyte transplantat, and humanized livestock liver t … More ransplant have gained great attention, as a new therapeutic approach and a new donor-source over severe situation of donor shortage. To across the xenogeneic barrier so that human cells could well fix and proliferate in porcine organ, even in the pig with immunological deficiency, it is necessary to understand the mechanism underlying the host-derived non-specific elimination system and to improve the proliferation ability of graft cells The purpose of this study, therefore, is to investigate how xenogeneic cellular grafts can fix and proliferate in porcine liver.Human-derived hepatic cell line, THLE5b, was used to evaluate its survival ability in mice liver. Adnovirus-based p21 transgene, a molecle known to stop the cell cycle, was transferred into the liver of CB17SCID mice. Partial hepatic resection was completed in the mice that induced p21 transgene over-expressed. Our data indicated that the potential of liver regeneration after partial resection falls down in p21 gene-transferred mice by evaluating the alteration of liver weight and the DNA synthesis. The THLE5b cells, developed as a cell source of human parenchymal hepatocye, proliferate well in vitro, but can only remain its normal morphologic features in the CB17SCID mice. The THLE5b cells, however, start proliferation in partial resected liver of p21 transgene mice. Our study demonstrate that p21 gene-transfer exhibits a negative effect in the liver regeneration after partial liver resection, and it could provide a suitable environment supporting xenogeneic cell proliferation. Less