Biochemical Engineering Institute, Saarland University, Campus A 1.5, 66123, Saarbrücken, Germany.
Adv Biochem Eng Biotechnol. 2012;127:109-32. doi: 10.1007/10_2011_99.
Reaction rates or metabolic fluxes reflect the integrated phenotype of genome, transcriptome and proteome interactions, including regulation at all levels of the cellular hierarchy. Different methods have been developed in the past to analyse intracellular fluxes. However, compartmentation of mammalian cells, varying utilisation of multiple substrates, reversibility of metabolite uptake and production, unbalanced growth behaviour and adaptation of cells to changing environment during cultivation are just some reasons that make metabolic flux analysis (MFA) in mammalian cell culture more challenging compared to microorganisms. In this article MFA using the metabolite balancing methodology and the advantages and disadvantages of (13)C MFA in mammalian cell systems are reviewed. Application examples of MFA in the optimisation of cell culture processes for the production of biopharmaceuticals are presented with a focus on the metabolism of the main industrial workhorse. Another area in which mammalian cell culture plays a key role is in medical and toxicological research. It is shown that MFA can be used to understand pathophysiological mechanisms and can assist in understanding effects of drugs or other compounds on cellular metabolism.
反应速率或代谢通量反映了基因组、转录组和蛋白质组相互作用的综合表型，包括在细胞层次的所有水平的调控。过去已经开发了不同的方法来分析细胞内通量。然而，哺乳动物细胞的区室化、多种底物的不同利用、代谢物摄取和产生的可逆性、细胞生长行为的不平衡以及细胞在培养过程中对环境变化的适应，使得与微生物相比，哺乳动物细胞培养中的代谢通量分析（MFA）更具挑战性。本文综述了使用代谢物平衡方法进行 MFA 的方法，以及（13）C-MFA 在哺乳动物细胞系统中的优缺点。本文还介绍了 MFA 在优化生物制药生产的细胞培养工艺方面的应用实例，重点关注主要工业工作载体的代谢。在医学和毒理学研究中，哺乳动物细胞培养也起着关键作用。研究表明，MFA 可用于了解病理生理机制，并有助于了解药物或其他化合物对细胞代谢的影响。
