HEPATOCYTES--TRANSPLANTATION, EXTRACORPOREAL SUPPORT

全器官移植仍然是临床上唯一有效的治疗方法 治疗急性肝功能衰竭和特殊的肝脏遗传缺陷 导致肝功能衰竭的功能。然而，有几个因素 这些限制了活体移植在治疗急性髓细胞白血病中的应用 肝功能不全。这些问题包括：捐赠者可获得性有限，无法 在急需的时候获得一个器官，生命的需要- 免疫抑制时间长，成本高。我们提议的工作的目的是 要使用分离的正常肝细胞执行以下操作： A.开发新的细胞移植方法。 B.开发生物人工肝(BAL)。 细胞移植。分离的肝细胞将主要用于治疗 特定的遗传性肝脏缺陷。在这些情况下，更正 肝功能缺陷会导致肝功能正常。使用的潜在优势 离体肝细胞移植包括：a.细胞冷冻保存；b. 体外细胞操作，以降低免疫原性或纠正 利用基因转移技术从单一捐赠者的细胞中获得遗传缺陷 用于治疗多个收件人，以及D.简单的技术 潜在的低成本。在过去的五年里，我们进行了研究 这导致了成功的腹膜腔内注射方法的发展 我们计划继续这些研究，特别是 解决这项技术的两个主要局限性：a.缺乏长期 功能和B.需要免疫抑制。此外，我们还将 特异性地研究肝细胞体外转染，检测肝细胞的命运和 转基因肝细胞在移植后和体内的稳定性 通过发展原位肝细胞转基因技术。 生物人工肝。BAL将主要用于治疗急性肝病 失败了。在过去的五年中，我们开发和测试了BAL系统 它利用微载体附着的正常肝细胞在外部 中空纤维生物反应器的表面。我们计划继续与 对这一制度的有效性和特点进行界定，以完善其 此外，还开发和测试了其他新型的BAL系统。这个 这些系统的主要局限性仍然是组织可用性不足。 因此，我们将致力于开发一种利用异种生物 肝细胞。

Whole organ transplantation remains the only clinically effective method of treating both acute liver failure an specific genetic defects of liver function which result in liver failure. However, there are several factors which limit the application of live transplantation in the treatment of liver insufficiency. These include: limited donor availability, inability to obtain an organ at the time when it is urgently needed, need for life- long immunosuppression and high cost. The purpose of our proposed work is to use isolated normal hepatocytes to: A. Develop Novel Methods of Cell Transplantation. B. Develop A Bioartificial Liver (BAL). Cell Transplantation. Isolated hepatocytes will be sued primarily to treat specific genetic liver defects. In these instances, correction of the defect results in normal liver function. Potential advantages of use of isolated hepatocyte transplantation include: a. cell cryopreservation, b. in vitro cell manipulation to either decrease immunogenicity or correct genetic defects using gene transfer technology cells from single donor can be used to treat multiple recipients, and d. simple technique at potentially low cost. In the past five years we have carried out studies which resulted in the development of successful methods of intraperitoneal transplantation and we plan to continue these studies and specifically address two major limitations of this technique: a. lack of long-term function and b. need for immunosuppression. In addition, we will specifically study hepatocyte transfection in vitro, examining the fate and stability of transfected hepatocytes following transplantation and in vivo by developing techniques of in situ hepatocyte transfection. Bioartificial Liver. The BAL will be used primarily to treat acute liver failure. In the past five years we have developed and tested a BAL system which utilizes microcarrier-attached normal hepatocytes on the outer surface of a hollow-fiber bioreactor. We plan to continue working with this system to define its effectiveness and characteristics, to improve its function and, in addition, develop and test other novel BAL systems. The major limitation of these systems remains inadequate tissue availability. We will thus work towards developing a BAL which utilizes xenogeneic hepatocytes.