生物型人工肝模型的建立及其在实验中的应用  

Construction of Biological Artificial Liver and Its Application in The Experimental Research

作　　者：郭恩玲[1] 孔静[1] 刘彬彬[1] 姜清泉 

机构地区：[1]中国医科大学附属盛京医院胆道外科,辽宁沈阳110021

出　　处：《中国普外基础与临床杂志》2015年第9期1137-1142,共6页Chinese Journal of Bases and Clinics In General Surgery

基　　金：国家自然科学基金资助项目(项目编号:81100313);辽宁省博士启动基金资助项目(项目编号:20111105)~~

摘　　要：目的总结各种生物型人工肝模型建立方法的优缺点,为将来各种肝脏代谢及疾病的研究提供理论依据及技术支持。方法搜索相关文献,总结肝细胞的取材、分离、培养条件及培养方法。结果肝细胞的分离方法中,机械分离法对肝细胞的取材不限,较易获得肝细胞,但对肝细胞的破坏较大,并且得到的细胞数量较少,存活率低。酶消化法对肝细胞的取材不限,其关键在于对酶浓度和消化时间的调节,仅限于涉及肝细胞功能、肝细胞对外界的反应等少部分研究。灌流消化法仅适用于动物,现应用较多,Ca2+、胶原酶种类及其灌流速度、p H值、缓冲液及插管方式都影响其效果。在培养的过程中根据实验需要选择不同的培养方法,再适当地在培养基中加入不同的添加剂。不同的生物反应器有着不同的优缺点及适用条件。结论对于供体要根据不同的实验目的选择不同的细胞来源;无论是在分离过程中还是在培养过程中,可根据具体情况来调整浓度、灌流时间、选择不同的培养基种类,以选择不同的生物反应器,但各种方法都存在着各种各样的不足,这些不足等待着更多的研究者去改善。Objective To provide theoretical and technological support for further study of liver metabolism and disease by comparing the advantages and disadvantages of various artificial liver models （biological）. Methods Literatures were searched and compared to summarize the requirement for liver donor, isolation, and culture of hepatocyte. Results In the separation method of hepatocyte, mechanical separation method had no requirement for liver donor, and was easy to acquire hepatocyte, while the acquired hepatocyte would be destructed severely, and the survival rate was low. On the other hand, the restriction of the digestion of the hepatocytes to the liver cell samples was unlimited, while the key of the enzyme digestion method was to regulate the balance between enzyme concentration and digestion time, which was limited to function researches of hepatocyte, and research about the responds of hepatocyte against outside, and other few researches. Perfusion digestion method had been widely applied for animal test. The Ca^2＋, collagenase and perfusion rate, pH value, buffer, and intubation method all play vital roles. During the cultivation, we needed to choose different methods according to several experiments, and add different additives in the appropriate medium. Different biological reactors had different advantages, disadvantages, and applicable conditions. Condusions The donor selection is based on various experimental purposes to harvest hepatocytes from different sources. Whether on the separation process or on the cultivation process, according to the specific circumstances, such as the concentration, perfusion time, and the choice of different kinds of culture medium, we can choose different kinds of bioreactors, but all kinds of methods are still remained with multiple insufficiencies, which require more researchers to improve.

关 键 词：生物型人工肝 肝细胞 分离 培养 

分 类 号：R318.14[医药卫生—生物医学工程]