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Abstract：
A fundamental variable in culture medium is its pH, which must be controlled by an appropriately formulated buffering regime, since biological processes are exquisitely sensitive to acid-base chemistry. Although awareness of the importance of pH is fostered early in the training of researchers, there are no consensus guidelines for best practice in managing pH in cell cultures, and reporting standards relating to pH are typically inadequate. Furthermore, many laboratories adopt bespoke approaches to controlling pH, some of which inadvertently produce artefacts that increase noise, compromise reproducibility or lead to the misinterpretation of data. Here, we use real-time measurements of medium pH and intracellular pH under live-cell culture conditions to describe the effects of various buffering regimes, including physiological CO2/HCO3- and non-volatile buffers (e.g. HEPES). We highlight those cases that result in poor control, non-intuitive outcomes and erroneous inferences. To improve data reproducibility, we propose guidelines for controlling pH in culture systems.
摘要：
培养基中的一个基本变量是其pH值，必须由适当制定的缓冲机制控制，因为生物过程对酸碱化学非常敏感。尽管在研究人员的培训初期就培养了人们对pH值重要性的认识，没有关于细胞培养中pH管理的最佳实践的共识指南，与pH相关的报告标准通常是不充分的。此外，许多实验室采用定制的方法来控制pH值，其中一些无意中产生了增加噪音的人工制品，影响可重复性或导致对数据的误解。这里,我们使用活细胞培养条件下的培养基pH和细胞内pH的实时测量来描述各种缓冲方案的影响，包括生理CO2/HCO3-和非挥发性缓冲液(例如HEPES)。我们强调那些导致控制不良的案例，非直觉的结果和错误的推论。为了提高数据的可重复性，我们提出了控制培养系统pH值的指南。
