新型中空纤维编织型生物人工肝反应器的初步构建  

Preliminary design of a novel bioreactor with woven hollow fibers

作　　者：王英杰[1] 郭海涛[1] 刘鸿凌[1] 刘俊[1] 王宇明[1] 

机构地区：[1]第三军医大学附属西南医院全军感染病研究所,重庆400038

出　　处：《第三军医大学学报》2003年第6期469-471,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 39970 2 14 ;30 0 2 70 0 1) ;全国优秀博士基金资助项目 ( 19994 7)

摘　　要：目的　为体外生物人工肝支持系统构建一种新型中空纤维编织型生物反应器 ,提高传统中空纤维生物反应器的性能。方法　采用中空纤维编织方法 ,制作三维立体型生物反应器 ,对反应器进行物质传输试验后 ,将分离的新生实验小型猪肝细胞接种于中空纤维支架上 ,用培养液循环式人工毛细管培养系统进行培养 ,检测生物反应器的生物功能 ,观察培养猪肝细胞的酶漏出量。结果　传输试验显示 ,中空纤维编织型反应器的半透膜能快速传输含人白蛋白和酚红的RPMI 164 0培养液。加入肝细胞并培养后 ,用SDS/PAGE可从生物反应器内检测出猪肝细胞分泌的白蛋白 ,生物反应器内的猪肝细胞有效地将氯化铵转换成尿素。反应器实验中 ,肝细胞仅释放了少量的ALT、LDH。结论　初步研制出一种新的、符合生物人工肝基本要求的实验型生物反应器 ,该反应器内培养的肝细胞保持较好的细胞活力 。Objective To design a novel 3 D network bioreactor of extracorporeal bioartificial liver support system with woven hollow fibers so as to improve the functions of traditional hollow fiber bioreactor. Methods 3 D network bioreactor was made of hollow fibers by a weave method. After the transmission test, the hepatocytes isolated from experimental suckling minipigs were seeded onto the hollow fiber scaffold of bioreactor and cultured with an artificial capillary cell culture system. Both the biological function and enzyme release of the bioreactor and hepatocytes were examined. Results Transmission test revealed that the woven hollow fibers could transmit the RPMI 1640 medium containing human albumin and phenol red at high speed. After the cells were seeded and cultured, porcine albumin could be estimated by SDS/PAGE in the culture media of the bioreactor. The bioreactor and porcine hepatocytes could transform ammonia chloride into urea effectively. During the experiment, only a little of ALT and LDH was released from the cultured hepatocytes into the medium. Conclusion A novel experimental bioreactor that meets the needs of bioartificial liver has been preliminarily designed. The porcine hepatocytes in the bioreactor can maintain viability and keep the functions of biosynthesis and biotransformation.

关 键 词：生物反应器 肝细胞 细胞培养 生物人工肝 

分 类 号：R318[医药卫生—生物医学工程] R318.14[医药卫生—基础医学]