通过摄氧量分析评估流体动力压力对禽类细胞系 DuckCelt®-T17 生长的影响

IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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引用次数: 0

摘要

由于细胞对搅拌或气泡破裂产生的流体动力应力非常敏感,因此影响了涉及动物细胞培养的生物工艺的放大。在这里,我们研究了最近的一种细胞系--DuckCelt®-T17 禽细胞(以前曾用于病毒疫苗的生产)在摇瓶和 3 L 生物反应器中经历的流体动力应力。细胞压力通过监测培养基中的溶解氧来评估,溶解氧取决于培养过程中的氧转移率(OTR)和氧吸收率(OUR)。此外,还测定了最大生长速率(µmax)和代谢物概况等经典参数。此外,还开发了一个动态模型,可预测营养物质消耗、代谢废物产生、存活细胞数和 OUR,并根据摇瓶中测量的数据进行了验证。在搅拌罐生物反应器(STBR)中进行的实验表明,OUR 取决于细胞生长阶段和搅拌条件。当平均剪切速率超过 70 s-1 和最大剪切速率超过 840 s-1 时,细胞在指数生长阶段(没有营养和氧气限制)的耗氧量会发生显著变化,这表明细胞对剪切非常敏感。OUR 是确定细胞稳健生长的流体动力学条件的合适工具。在 STBR 中进行 DuckCelt®-T17 细胞培养的放大标准是剪切力和/或尖端速度。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Oxygen uptake rate analysis to evaluate the impact of hydrodynamic stress on the growth of the avian cell line DuckCelt®-T17
Scale-up of bioprocesses involving animal cell culture is hampered by the sensitivity of the cells to hydrodynamic stress, either from agitation or bubble bursting. Here, the hydrodynamic stress experienced by a recent cell line, the DuckCelt®-T17 avian cells, previously used for viral vaccine production, is investigated in shake flasks and in a 3 L bioreactor. Cell stress was assessed by monitoring the dissolved oxygen in the culture medium, which depends on Oxygen Transfer Rate (OTR) and Oxygen Uptake Rate (OUR) during cultivation. Classical parameters such as the maximum growth rate (µmax) and metabolite profiles were also determined. A dynamic model able to predict nutrient consumption, metabolic waste production, viable cell number and OUR was also developed and validated from the data measured in shake flasks. The experiments performed in the stirred tank bioreactor (STBR) show that OUR depended on both the cell growth phase and the stirring conditions. The oxygen consumption of the cells during the exponential growth phase (where there were no nutrient and O2 limitations) was significantly altered at average and maximum shear rates above 70 and 840 s−1, respectively, indicating highly shear-sensitive cells. OUR is a suitable tool to identify the hydrodynamic conditions for robust cell growth. The scale-up criteria to be favored for the DuckCelt®-T17 cell culture in STBRs would be the shear and/or the tip’s speed.
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来源期刊
Biochemical Engineering Journal
Biochemical Engineering Journal 工程技术-工程:化工
CiteScore
7.10
自引率
5.10%
发文量
380
审稿时长
34 days
期刊介绍: The Biochemical Engineering Journal aims to promote progress in the crucial chemical engineering aspects of the development of biological processes associated with everything from raw materials preparation to product recovery relevant to industries as diverse as medical/healthcare, industrial biotechnology, and environmental biotechnology. The Journal welcomes full length original research papers, short communications, and review papers* in the following research fields: Biocatalysis (enzyme or microbial) and biotransformations, including immobilized biocatalyst preparation and kinetics Biosensors and Biodevices including biofabrication and novel fuel cell development Bioseparations including scale-up and protein refolding/renaturation Environmental Bioengineering including bioconversion, bioremediation, and microbial fuel cells Bioreactor Systems including characterization, optimization and scale-up Bioresources and Biorefinery Engineering including biomass conversion, biofuels, bioenergy, and optimization Industrial Biotechnology including specialty chemicals, platform chemicals and neutraceuticals Biomaterials and Tissue Engineering including bioartificial organs, cell encapsulation, and controlled release Cell Culture Engineering (plant, animal or insect cells) including viral vectors, monoclonal antibodies, recombinant proteins, vaccines, and secondary metabolites Cell Therapies and Stem Cells including pluripotent, mesenchymal and hematopoietic stem cells; immunotherapies; tissue-specific differentiation; and cryopreservation Metabolic Engineering, Systems and Synthetic Biology including OMICS, bioinformatics, in silico biology, and metabolic flux analysis Protein Engineering including enzyme engineering and directed evolution.
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