HYBRID ARTIFICIAL LIVER USING IMMOBILIZED HEPATOCYTES

目的是固定肝细胞用于人工肝， 肝脏装置 将采用已建立的技术分离肝细胞， 胶原酶消化并固定在海藻酸钠凝胶基质中。 这种藻酸盐凝胶将提供机械保护的固定 细胞 在这项工作中开发的原则和技术将提供一个 急性和慢性疾病治疗模式设计的合理依据 肝衰竭 肝细胞的活力和功能将被 从短期和长期来看。 可行性将使用 用台盼蓝和荧光染料进行活体染料测定。 功能 将通过测量以下反应动力学来评估肝细胞的 混合功能氧化酶系统（细胞色素P450），尿素循环，和 结合酶（UDP-葡糖醛酸转移酶和硫酸转移酶）。 在急性肝衰竭动物模型中，将使用固定化细胞 无论是腹膜内移植还是体外循环， 提供短期肝脏支持。 在过去的9年中，申请人已经完成了5年的普通外科手术 马萨诸塞州总医院的住院医师培训计划， 获得化学工程硕士和博士学位， 马萨诸塞州理工学院。 他将把教育应用于 工程和他的临床训练，外科的发展， 人工肝支持技术。 虽然申请人有 研究背景，这项研究是在工程，而不是在 生物化学、细胞生物学和消化疾病生理学。 的 申请人将需要进一步的培训和发展，在学习这些 成为肝脏独立研究者的基本科学技能 research. 申请人的赞助商将是Kurt Isselbacher博士， 马萨诸塞州总医院的医疗服务。 伊塞尔巴赫博士 将提供必要的环境，以发展基本的科学技能， 肝生化 马萨诸塞州总医院外科服务处 医院和外科，哈佛医学院，将 提供学术承诺和实验室空间。 保罗·罗素博士 来自马萨诸塞州总医院外科的医生 在项目的移植和免疫方面进行合作。

The objective is the immobilization of hepatocytes for use in an artificial liver device. Hepatocytes will be isolated by established techniques using collagenase digestion and immobilized in a sodium alginate gel matrix. This alginate gel will provide mechanical protection of the immobilized cells. The principles and technology developed in this work will provide a rational basis for design of treatment modalities for acute and chronic liver failure. The viability and function of the hepatocytes will be examined in the short and long term. The viability will be assessed using vital dye assays with both trypan blue and fluorescence dyes. The function of the hepatocytes will be assessed by measuring the reaction kinetics for the mixed function oxidase system (cytochrome P450), the urea cycle, and conjugating enzymes (UDP-glucuronyltransferase and sulfate transferase). In acute liver failure animal models, the immobilized cells will be used both as an intraperitoneal transplant and in an extracorporeal circuit to provide short term liver support. In the last 9 years, the applicant has completed the 5 year General Surgery residency training program at the Massachusetts General Hospital and also received a masters and a doctoral degree in Chemical Engineering at the Massachusetts Institute of Technology. He will apply the education in engineering and his clinical training in Surgery to the development of techniques for artificial liver support. Though the applicant has a background in research, this research was in engineering and not in biochemistry, cell biology, and physiology of digestive diseases. The applicant will require further training and development in learning these basic science skills to become an independent investigator in liver research. The sponsor of the applicant will be Dr. Kurt Isselbacher from the Medical Service of the Massachusetts General Hospital. Dr. Isselbacher will provide the environment necessary to develop basic science skills in liver biochemistry. The Surgical Service of the Massachusetts General Hospital and the Department of Surgery, Harvard Medical School, will provide the academic commitment and laboratory space. Dr. Paul Russell from the Surgical Service of the Massachusetts General Hospital will collaborate on transplant and immunological aspects of the project.