WAVETM反应器和搅拌瓶中微载体球转球放大培养的比较

目的:哺乳动物细胞目前已广泛用于生物工程药物如单抗和疫苗的生产.而用于贴壁细胞规模化培养的微载体,也应时应需得以开发并应用于生物制药.贴壁细胞微载体培养在搅拌罐和WAVETM反应器中都能进行.而如要进行进一步的放大培养,球转球工艺不可或缺.为了发展球转球这一新的放大技术,以及考量WAVETM反应器这种新型大规模培养设备的应用性,大量的细胞培养和球转球实验在WAVETM反应器和搅拌瓶中进行.收集到的数据得以分析比较.方法:将Vero细胞分别接入WAVETM反应器和搅拌瓶中用微载体Cytodex 1进行培养.适当补充营养并控制温度、pH等培养条件使细胞增殖.长满微载体的细胞用清洗、消化等球转球工艺的一系列步骤而分离,并放大接种到新的培养体系.球转球工艺的有效性通过记录并统计分析细胞消化分离的回收率,以及细胞重新接种生长的存活力来评估.结果:统计学分析比较WAVETM反应器和搅拌瓶中得到的细胞分离回收率分别是67.56％和39.39％,数理统计P值小于0.0003;细胞重新接种存活率分别是95.17％和78.45％,P值等于0.0107.结论:在WAVETM反应器中进行的球转球放大工艺,其总体表现和有效性远高于在搅拌瓶中得到的结果.在WAVETM反应器中培养的Vero细胞有很好的细胞状态,作为种子链和生产用罐相比搅拌型反应罐均有很大的优越性.

WAVETM Bioreactors and Spinner Flasks:Comparison of the Process Performance of Microcarrier Bead to Bead Transfer and Scale up

Objective: Mammalian cell lines are widely used in biopharmaceuticals,such as the manufacturing of monoclonal antibodies,recombinant proteins and vaccines.For large scale adherent cell culture,microcarriers are developed and widely used in biopharmaceuticals.Adherent cell culture on microcarriers andbe done in both WAVETM reactors and stir tanks.However,to successfully scale up the microcarrier cultures,bead to bead transfer process is necessary.To develop bead to bead transfer,the new scale up technique,and evaluate the reliability of WAVETM reactor,a new generation of large scale fermenters,plenty of cell culture and bead to bead transfer experiments were done in WAVETM reactors and spinner flasks.The data were collected and analyzed.Method: Vero cells were inoculated and cultured with microcarrier Cytodex 1 in WAVETM reactors and spinner flasks,respectively.For good cell propagation,the culture conditions such as pH,temperature and nutrients were controlled appropriately.The cells grown confluent on the microcarriers were separated through bead to bead transfer process including wash,trypsin digesting,and re-inoculating,to be transferred a new scaled up culture.The efficiencies of bead to bead transfer process were evaluated through recording and analyzing the cell recoveries during wash and digestion,and cell growth viabilities after re-inoculum.Results: Statistical analysis showed that the average of cell recoveries of the bead to bead transfer processes from WAVETM reactors and spinner flasks were 67.56% and 39.39%,respectively.P value was below 0.000 3.The average inoculums re-attach efficiency from WAVETM reactors and spinner flasks were 95.17% and 78.45%,respectively.P value was 0.010 7.Conclusion: Bead to bead transfer process done in WAVETM reactors got much better performance in terms of cell recovery and cell viability,comparing with that in spinner flasks.Vero cells grown in WAVETM are in good conditions.As either seed train or production tank,WAVETM reactors wound be muchsuperior comparing with stir tanks.