Modulation of Pancreatic Cancer Cell Activity in CO2-Free Culture Using TES and MOPS Buffers

Abstract

The study aim was to test whether Good's buffers (TES, MOPS) can stabilize pH, improve assay consistency and enhance reliability in CO₂-independent conditions.

In Phase I, both TES and MOPS buffers were found to introduce substantial variability into the assays. This volatility extended beyond treated samples, as even control groups occasionally produced inconsistent results without any intervention. While frustrating, these findings are valuable they emphasize the need for caution when interpreting MTT or LDH assays, particularly in co-culture systems with bacterial infection. This is especially true when the LDH assay is applied post-lysis, where results may be further confounded.
In Phase II, the dataset was limited (two biological repeats, no technical repeats), and thus no firm conclusions can be drawn. Preliminary trends suggest that passage number may influence outcomes: at higher passages, a gap appeared between CO₂ and CO₂-free controls, which could be partially equilibrated by buffer supplementation, until higher concentrations introduced toxicity, At lower passages, however, this baseline gap was absent, and buffer addition instead led to erratic behaviour with no consistent pattern.
Taken together, the results highlight both the challenges and the nuances of working with PANC-1 cells under altered buffering systems, If there is a single take-home message from Phase I, it is this: be wary of quick interpretations, for these cells are clearly mischievous