Cell Technology Group, National Institute for Bioprocessing Research and Training (NIBRT), Blackrock, Co., Dublin, Ireland.
School of Chemical and Bioprocess Engineering, University College Dublin (UCD), Blackrock, Co., Dublin, Ireland.
Biotechnol Prog. 2024 Sep-Oct;40(5):e3469. doi: 10.1002/btpr.3469. Epub 2024 Apr 13.
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.
可靠的监测生物反应器中的哺乳动物细胞对于生物制药生产至关重要。台盼蓝排斥是一种用于确定细胞密度和活力的方法,已经使用了一百多年来监测培养中的细胞,是生物制造中的当前标准方法。然而,这种方法有许多缺点,并且人们越来越需要更详细和在线测量生物反应器中细胞的生长。本文评估了一种用于单细胞分析的新型动态成像系统。这些数据表明,成像系统生成的可比总细胞密度、活细胞密度和存活率数据与传统的台盼蓝计数方法非常吻合,适用于具有工业相关性的中国仓鼠卵巢 (CHO) 细胞系。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在 6 天的生物反应器培养过程中监测哺乳动物细胞健康状况的趋势。该系统还能够进行在线取样,无需离线取样,并能够实时监测生物反应器培养中的细胞。