摘要
目的：探讨大量培养高活性人肝细胞的理想方法，为肝脏病研究提供良好的细胞材料。方法：在限制贴壁、定时振荡条件下，采用载体ｃｙｔｏｄｅｘ３为材料进行人肝细胞的微载体培养。结果：预先将体外两步灌流分离的人肝细胞振荡３０分钟后，肝细胞易与微载体相粘附．在被覆聚羟乙基异丁烯酸的培养瓶中继续间歇性振荡２４小时，约７０％的肝细胞附着于微载体之上。在使用激素限定条件培养液培养的１５天内，肝细胞保持蛋白分泌功能和多细胞粘附聚集球形体形态特征。结论：本法培养人肝细胞具有细胞数量多、密度大和活性好的特点，适宜于体外生物人工肝支持系统、肝细胞移植以及培养肝细胞因子的提取。
Objective: To explore an optimal method for large-scale cultivation of human hepatocytes and to provide a good material for the study of liver disease. Methods: Culture of human hepatocytes on microcarriers was carried out by cytodex 3 on the conditions of the cells restricted to attached dish and the media was stirred at regular intervals. Results: Human hepatocytes obtained from extracorporeal two-stage perfusion method were efficiently adhered on microcarriers when the isolated cells has been stirred after 30min. The adherence rate of hepatocytes on microcarriers was about 70% on the flasks coated with poly(2-hydroxythl methacrylate) after 24 hours of incubation and stirring at regular interval. In hormonally defined media, human hepatocytes on microcarriers retained the ability to secrete proteins and had a multicellular adherent and aggregated apheroidal characteristics up to 15th day. Conclusions: This method developed for large-scale and high-density production of hepatocytes on microcarriers will allow one to obtain active human liver cells suitable for extracorporeal bioartificial liver support systems、hepatocytic transplantation and hepatocytic cytokine extraction.
出处
《中华消化杂志》
CAS
CSCD
北大核心
1997年第4期191-194,共4页
Chinese Journal of Digestion
基金
国家自然科学基金
关键词
细胞培养
肝细胞
人
载体
Cell culture Hepatocytes Human Carrier
