支持高密度BHK-21细胞培养和高产FMD病毒的无血清培养基开发与优化  

Development and Optimization of Serum-Free Medium for Highdensity Culture of Suspended BHK-21 Cells and High-yield of FMD Virus

作　　者：陈敏 刘旭平 赵亮[2] CHEN Min;LIU Xu-ping;ZHAO Liang(Shanghai Bioengine Biotechnology Co.Ltd.,Shanghai 201203;State Key Laboratory of Bioreacter Engineering,East China University of Science and Technology,Shanghai 200237)

机构地区：[1]上海倍谙基生物科技有限公司,上海201203 [2]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《生物技术通报》2020年第10期62-71,共10页Biotechnology Bulletin

摘　　要：优化无血清培养基,以实现BHK-21悬浮细胞的超高密度生长和口蹄疫病毒高效扩增。依据BHK-21细胞的代谢特点和动力学分析识别无血清培养基中的关键营养成分,优化组分浓度,对比优化前后细胞的生长和产毒能力,并在生物反应器中验证。优化后的无血清培养基支持批培养模式下最高活细胞密度达到1.78×107 cells/mL,代谢副产物积累的情况得到改善;口蹄疫病毒TCID50与含低比例血清的培养基相比具有相似水平,约为7.25 lgTCID50/0.1mL;146s含量与低血清培养基相比提高50.7%,达到15.6μg/mL。以细胞的生长需求和代谢规律为基础,优化营养成分和降低代谢副产物积累能有效解除“细胞密度效应”,实现超高细胞密度接毒和高产口蹄疫病毒。Based on the high-density suspension culture of BHK-21 cells,this study is to optimize the serum-free medium in order to improve proliferation of the BHK-21 cells and amplification of foot-and-mouth virus(FMDV).Firstly,the key nutrients in the serum-free medium were determined based on the metabolic characteristics and dynamics analysis of the BHK-21 cells.Then,the composition of the medium was optimized.Finally,the growth and FMDV-producing ability of the BHK-21 cells between before and after the optimization were compared and verified in bio-reactor.The maximum viable cell density reached 1.78×107 cells/mL during batch culturing of the BHK-21 cells in optimized serum-free medium(Opt-SFM),and the situation of accumulating metabolic by-products was improved.Compared with nonoptimized medium containing 1%(V/V)serum,TCID50 of the produced FMDV in the Opt-SFM showed a similar level at 7.25 lgTCID50/0.1 mL,while the yield of 146s particals increased by 50.7%to 15.6 g/mL.In conclusion,based on the requirements of cell growth and metabolic characteristics,optimizing nutrients and reducing metabolic by-products accumulation may effectively eliminate“cell density effect”and achieve ultra-high cell density and high yield of foot-and-mouth virus.

关 键 词：BHK-21悬浮细胞 无血清培养基 高细胞密度 高产口蹄疫病毒 

分 类 号：S855.3[农业科学—临床兽医学]