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Studies on Biomimetic Artificial Liver

仿生人工肝的研究

基本信息

批准号：
63570585
负责人：
KASAI Shinichi
金额：
$ 1.28万
依托单位：
Asahikawa Medical College
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

项目摘要

We have developed a prototype of hybrid artificial liver support system using isolated hepatocytes and have reported previously. In those systems, the viability of hepatocytes could not be maintained over ten hours because of a floating system. The purpose of this study is to evaluate the usefulness of hepatocyte culture substrata to maintain the viability and functions of hepatocytes for longer time developing a hybrid artificial liver.Male Wister rats weighing about 2OOg were used to obtain isolated hepatocytes by collagenase digestive method. As natural substrata, collagen, fibronectin, vitronectin and biomatrix were examined. Biomatrix was prepared from dog liver by the method of Rojkind. As synthetic substrata, lactose-carrying polystyrene (PVLA) and microcarrier Cytodex 3 were used, and entrapped hepatocytes within carcium arginate gel was also examined.Cell attachment of isolated hepatocytes and their behavior were observed microscopically on the dishes coated with collagen, fibronectin, vitronectin or biomatrix, PVLA coated dish and Cytodex 3 were also investigated for the hepaticytes attachment. Metabolic functions were examined in dish culture and in spinner flask incubations.Hepatocytes on the dish coated with substrata such as collagen, fibronectin, vitronectin attached and spread well 24 hours after seeding, but hepatocytes did not spread on biomatrix. On the dish with PVLA, hepatocytes attached well but not spread, and the hepatocytes also attached on Cytodex 3. In dish culture test and spinner flask test, remarkable decrease of NH_3 concentrations with a little increase of BUN and glucose concentrations were observed in each materials.

我们已经开发了一个原型的混合人工肝支持系统使用分离的肝细胞和以前的报告。在这些系统中，由于漂浮系统，肝细胞的活力不能维持超过10小时。为探讨肝细胞培养基质对维持肝细胞活力和功能的有效性，以200g左右的雄性Wister大鼠为实验动物，采用胶原酶消化法分离肝细胞。作为天然基质，胶原蛋白，纤维连接蛋白，玻连蛋白和生物基质进行了检查。采用Rojkind法从狗肝中制备生物基质。采用载乳糖聚苯乙烯（PVLA）和微载体Cytodex 3作为合成基质，并检查了arginate凝胶中包埋的肝细胞，在显微镜下观察了分离的肝细胞在胶原蛋白、纤连蛋白、玻连蛋白或生物基质包被的培养皿、PVLA包被的培养皿和Cytodex 3上的细胞附着及其行为。在平皿培养和转瓶培养中检测了肝细胞的代谢功能，接种后24小时，肝细胞在胶原、纤维连接蛋白、玻连蛋白等基质包被的平皿上贴壁铺展良好，但肝细胞在生物基质上没有铺展。在含有PVLA的培养皿上，肝细胞附着良好但未扩散，肝细胞也附着在Cytodex 3上。在平皿培养试验和转瓶试验中，每种材料的NH_3浓度均显著降低，而BUN和葡萄糖浓度略有升高。