Liu Chi-Hsien, Liu Yi-Xin, Wu Wei-Chi
Department of Chemical and Materials Engineering, Chang Gung University, 259, Wen-Hwa First Road, Kwei-Shan, Taoyuan, 333, Taiwan.
Research Center for Chinese Herbal Medicine and Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, 261, Wen-Hwa First Road, Taoyuan, Taiwan.
Cytotechnology. 2018 Dec;70(6):1631-1642. doi: 10.1007/s10616-018-0255-z. Epub 2018 Oct 3.
Most bio-industrial mammalian cells are cultured in serum-free media to achieve advantages, such as batch consistency, suspended growth, and simplified purification. The successful development of a serum-free medium could contribute to a reduction in the experimental variation, enhance cell productivity, and facilitate biopharmaceuticals production using the cell culture process. Commercial serum-free media are also becoming more and more popular. However, the cell line secrets its own recombinant product and has special nutritional requirements. How can the composition of the proprietary medium be adjusted to support the specific cell's metabolism and recombinant protein? This article uses statistical strategies to modify the commercial medium. A design of experiments is adopted to optimize the medium composition for the hybridoma cell in a serum-free condition. The supplements of peptone, ferric citrate, and trace elements were chosen to study their impact on hybridoma growth and antibody production using the response surface methodology. The stimulatory effect of the developed formulation on hybridoma growth was confirmed by the steepest ascent path. The optimal medium stimulated the hybridoma growth and antibody production in three diverse systems: a static plate, an agitated spinner flask, and a hollow fiber reactor. The cells in the developed serum-free medium had a better antibody production as compared to that in the commercial medium in the hollow fiber reactor. Our results demonstrated that the facile optimization for medium and antibody production was successfully accomplished in the hybridoma cells.
大多数生物工业哺乳动物细胞在无血清培养基中培养，以获得诸如批次一致性、悬浮生长和简化纯化等优势。无血清培养基的成功开发有助于减少实验变异、提高细胞生产力，并促进使用细胞培养工艺生产生物制药。商业无血清培养基也越来越受欢迎。然而，细胞系会分泌自身的重组产物，并有特殊的营养需求。如何调整专有培养基的成分以支持特定细胞的代谢和重组蛋白？本文采用统计策略对商业培养基进行改良。采用实验设计在无血清条件下优化杂交瘤细胞的培养基成分。选择蛋白胨、柠檬酸铁和微量元素的添加物，使用响应面法研究它们对杂交瘤生长和抗体产生的影响。通过最速上升路径证实了所开发配方对杂交瘤生长的刺激作用。优化后的培养基在三种不同系统中刺激了杂交瘤生长和抗体产生：静态平板、搅拌转瓶和中空纤维反应器。在中空纤维反应器中，与商业培养基相比，在开发的无血清培养基中的细胞具有更好的抗体产生能力。我们的结果表明，在杂交瘤细胞中成功实现了培养基和抗体产生的简便优化。
