HYBRID ARTIFICIAL LIVER USING IMMOBILIZED HEPATOCYTES

使用固定化肝细胞的混合人工肝

• 批准号: 3080591
• 负责人: RONALD GARY TOMPKINS
• 金额: $ 7.56万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-01-01 至 1991-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3080591
• 关键词: cytochrome P450; histology; liver; liver cells; radiotracer; tissue /cell culture; transplantation; urea cycle

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.

目的是固定肝细胞以用于人工 肝脏装置。 将通过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。 这种藻酸盐凝胶将为固定化物提供机械保护 细胞。 这项工作中开发的原理和技术将提供 急性和慢性治疗方式设计的合理依据 肝功能衰竭。 肝细胞的活力和功能将 从短期和长期来看。 将使用以下方法评估可行性 使用台盼蓝和荧光染料进行活体染料测定。 功能 将通过测量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-葡萄糖醛酸转移酶和硫酸盐转移酶）。 在急性肝衰竭动物模型中，将使用固定化细胞 作为腹膜内移植和体外循环 提供短期肝脏支持。 在过去 9 年内，申请人已完成 5 年普通外科课程 马萨诸塞州总医院的住院医师培训计划以及 获得化学工程硕士和博士学位 麻省理工学院。 他将把所受的教育应用到 工程和他在外科方面的临床训练，以发展 人工肝支持技术。 尽管申请人有 研究背景，这项研究是工程领域的，而不是 生物化学、细胞生物学和消化系统疾病生理学。 这 申请人将需要进一步的培训和发展来学习这些 成为肝脏独立研究者的基本科学技能 研究。 申请人的担保人将是来自德国的 Kurt Isselbacher 博士 马萨诸塞州总医院的医疗服务。 伊塞尔巴赫博士 将为发展基本科学技能提供必要的环境 肝脏生化。 马萨诸塞州总医院的外科服务 哈佛医学院医院和外科部将 提供学术承诺和实验室空间。 保罗·拉塞尔博士 来自马萨诸塞州总医院外科服务中心的 就该项目的移植和免疫学方面进行合作。