HYBRID ARTIFICIAL LIVER USING IMMOBILIZED HEPATOCYTES

目的是将肝细胞固定化，以用于人工 肝脏装置。肝细胞将通过现有的技术分离出来，使用 胶原酶消化并固定在海藻酸钠凝胶基质中。 这种海藻酸凝胶将为固定化的 细胞。这项工作中开发的原则和技术将提供一种 急慢性疾病治疗模式设计的合理依据 肝功能衰竭。肝细胞的活性和功能将是 从短期和长期两方面进行考察。可行性将通过以下方式评估 用台盼蓝和荧光染料对活性染料进行检测。该功能 将通过测量肝细胞的反应动力学来评估 混合功能氧化酶系统(细胞色素P450)，尿素循环，以及 结合酶(UDP-葡萄糖醛酸基转移酶和硫酸盐转移酶)。 在急性肝功能衰竭动物模型中，固定化细胞将用于 无论是作为腹膜内移植还是体外循环 提供短期的肝脏支持。 在过去的9年里，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 在伦敦大学获得化学工程硕士和博士学位 麻省理工学院。他将把这项教育应用于 工程学和他在外科的临床培训对发展 人工肝支持技术。虽然申请者有一个 研究背景，这项研究是在工程领域，而不是在 消化疾病的生物化学、细胞生物学和生理学。这个 申请人将需要在学习这些方面进行进一步的培训和发展 成为肝脏独立研究人员的基本科学技能 研究。申请人的赞助人将是库尔特·伊斯尔巴赫博士 马萨诸塞州总医院的医疗服务。伊塞尔巴赫博士 将提供发展基本科学技能所需的环境 肝脏生化。马萨诸塞州总医院的外科服务 医院和哈佛医学院外科将 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服务部的威尔 就移植和免疫学方面的项目进行合作。

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.