Single cell analysis of Chinese hamster ovary cells during a bioprocess using a novel dynamic imaging system

IF 2.5 3区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology Progress Pub Date : 2024-04-13 DOI:10.1002/btpr.3469
Laura Breen, James Flynn, Adam Bergin, Evangelia Flampouri, Michael Butler
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Abstract

Reliable monitoring of mammalian cells in bioreactors is essential to biopharmaceutical production. Trypan blue exclusion is a method of determining cell density and viability that has been used for over one hundred years to monitor cells in culture and is the current standard method in biomanufacturing. This method has many disadvantages however and there is a growing demand for more detailed and in-line measurements of cell growth in bioreactors. This article assesses a novel dynamic imaging system for single cell analysis. This data shows that comparable total cell density, viable cell density and percentage viability data shown here, generated by the imaging system, aligned well with conventional trypan blue counting methods for an industrially relevant Chinese Hamster Ovary (CHO) cell line. Furthermore, detailed statistical analysis shows that the classification system used by the PharmaFlow system can reveal trends of interest in monitoring the health of mammalian cells over a 6-day bioreactor culture. The system is also capable of sampling at-line, removing the necessity for taking samples off-line and enabling real time monitoring of cells in a bioreactor culture.

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利用新型动态成像系统对生物加工过程中的中国仓鼠卵巢细胞进行单细胞分析
可靠地监测生物反应器中的哺乳动物细胞对生物制药生产至关重要。胰蓝排除法是一种确定细胞密度和存活率的方法,用于监测培养中的细胞已有一百多年的历史,也是目前生物制造中的标准方法。但这种方法有许多缺点,因此对生物反应器中细胞生长的更详细和在线测量的需求日益增长。本文评估了一种用于单细胞分析的新型动态成像系统。数据显示,对于与工业相关的中国仓鼠卵巢(CHO)细胞系,成像系统生成的可比总细胞密度、存活细胞密度和存活率数据与传统的胰蓝计数法十分吻合。此外,详细的统计分析表明,PharmaFlow 系统使用的分类系统可以揭示监测哺乳动物细胞在 6 天生物反应器培养过程中健康状况的相关趋势。该系统还能在线取样,无需离线取样,实现了对生物反应器培养细胞的实时监测。
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来源期刊
Biotechnology Progress
Biotechnology Progress 工程技术-生物工程与应用微生物
CiteScore
6.50
自引率
3.40%
发文量
83
审稿时长
4 months
期刊介绍: Biotechnology Progress , an official, bimonthly publication of the American Institute of Chemical Engineers and its technological community, the Society for Biological Engineering, features peer-reviewed research articles, reviews, and descriptions of emerging techniques for the development and design of new processes, products, and devices for the biotechnology, biopharmaceutical and bioprocess industries. Widespread interest includes application of biological and engineering principles in fields such as applied cellular physiology and metabolic engineering, biocatalysis and bioreactor design, bioseparations and downstream processing, cell culture and tissue engineering, biosensors and process control, bioinformatics and systems biology, biomaterials and artificial organs, stem cell biology and genetics, and plant biology and food science. Manuscripts concerning the design of related processes, products, or devices are also encouraged. Four types of manuscripts are printed in the Journal: Research Papers, Topical or Review Papers, Letters to the Editor, and R & D Notes.
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Non-thermal plasma decontamination of microbes: a state of the art. Mechanistic model of minute virus of mice elution behavior in anion exchange chromatography purification. Comparing in silico flowsheet optimization strategies in biopharmaceutical downstream processes. General strategies for IgG-like bispecific antibody purification. Issue Information
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