HepG2细胞滚瓶培养工艺优化及硒酸壳聚糖对其抑制作用  

Optimization of Incubation of HepG2 Cells in Roller Bottle and Apoptosis Induced by Seleno-Chitosan

作　　者：纪海玉 冯莹莹 于娟[1] 彭雪丽 董晓丹 刘安军[1] 

机构地区：[1]天津科技大学食品工程与生物技术学院,天津300457

出　　处：《食品研究与开发》2018年第3期189-195,共7页Food Research and Development

基　　金：省部级农业部项目“低盐低硝腌腊肉制品生产关键技术装备研究与示范”(201303082-3)

摘　　要：以HepG2为研究对象,细胞产量为考察指标,在单因素试验的基础上,采用响应面分析法,优化了HepG2细胞滚瓶培养工艺:接种量4×10~7个/瓶,培养时间48 h,滚瓶转速9 r/min,此条件下每滚瓶HepG2细胞产量预测值10.56×10~7个/瓶,验证试验得到实际细胞量为10.88×10~7个/瓶,与理论预测值相比,其相对误差约为3.03%。体外抗肿瘤试验表明,硒酸壳聚糖(Seleno-Chitosan,CS)对96孔板和滚瓶培养HepG2细胞抑制作用IC_(50)值分别为169μg/m L和245μg/m L,表明细胞大量聚集可产生药物拮抗现象。该试验结果为抗肿瘤疫苗研发及肿瘤药物治疗提供理论依据。Based on single-factor experiments, response surface methodology was employed to optimize the cultivation process of HepG2 cellsin roller bottles. The optimal culture conditions were found that 4×107 HepG2 cells were incubated each bottle cultured for 48 h with rotate speed of 9 r/min.The predicted HepG2 cells yield under these conditions was 10.56×107 cells/bottle and experimental value was 10.88×107 cells/bottle with the relative error being about 3.03 %.The results in vitro antitumor experiments showed that seleno-chitosan could significantly inhibit the proliferation of HepG2 cells in 96-well plate and roller bottles with IC50 of 169 μg/mL and 245 μg/mL respectively, suggesting that the HepG2 cells cultured in roller bottles would cause cells aggregation and generate drug antagonists. The experimental results provide a theoretical basis for development of cancer vaccine and drug treatment of tumor.

关 键 词：HEPG2细胞 滚瓶培养 硒酸壳聚糖 抗肿瘤 

分 类 号：Q813[生物学—生物工程] R73-36[医药卫生—肿瘤]