Mixture与响应面法结合开发BHK-21细胞无血清悬浮培养基  

Optimization of Suspended Serum-free Medium for BHK-21 Cells by Combining Mixture with Response Surface Analysis

作　　者：汪梁[1] 刘旭平[1] 谭文松[1] 

机构地区：[1]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》2015年第9期70-78,共9页Biotechnology Bulletin

基　　金：国家自然科学基金项目(21206040;21406066);国家"863"计划项目(2012AA02A303);国家重大专项(2013ZX10004003-003-003);中央高校基本科研业务费(WF1214035)

摘　　要：采用Mixture与响应面实验设计相结合的方法开发BHK-21细胞无血清悬浮培养基。在实验室已知配方的6种培养基A-F的基础上,通过Mixture实验筛选出BHK-21细胞无血清培养基的最优组合为A∶B∶C∶D∶E∶F=0∶0∶11∶0∶9∶0。利用响应面法针对培养基中的几种关键组分进行浓度优化,确定谷氨酰胺、酪氨酸、牛血清白蛋白和钙离子的最优浓度分别为3 mmol/L、2.5 g/L、0 g/L和0 mmol/L。该无血清悬浮培养基能很好地支持BHK细胞悬浮生长,培养时细胞最大活细胞密度可达140.21×105个/m L,比商业培养基提高了1.95倍。采用Mixture试验设计和响应面分析法能够在较短的时间内开发出适合BHK-21细胞生长的无血清悬浮培养基,为采用BHK-21细胞大规模工业化生产口蹄疫疫苗奠定了基础。This study is to optimize the suspended serum-free medium for BHK-21 cells by combining Mixture experiment design with response surface analysis（RSA）. Based on 6 media A-F of known formulation in the laboratory, the optimal combination of the medium with A：B：C：D：E：F=0：0：11：0：9：0 was screened by the Mixture test. The contents of several key constituents in the medium were optimized by RSA, and the optimal concentrations of glutamine, tyrosine, bovine serum albumin（BSA）and calcium ion were 3 mmol/L, 2.5 g/L, 0 g/L and 0 mmol/L respectively. The serum-free medium effectively supported the suspending growth of BHK cells. The maximum viable cell density of BHK cells in cell suspension culture reached 140.21×105 cells/ml, which was 1.95 times in basal medium. The suspended serum-free medium for BHK-21 cells was developed by combining Mixture experiment design with RSA, which lays a foundation for the industrial production of foot-and-mouth disease（FMD）vaccine while using BHK-21 cells.

关 键 词：BHK-21细胞 无血清悬浮培养基 Mixture实验设计 响应面分析 

分 类 号：S859.797[农业科学—临床兽医学]