Figure 5
From: Rational cell culture optimization enhances experimental reproducibility in cancer cells
Live cell numbers, concentrations of various metabolites, and pH in fresh and spent media extracts 24 hours after treatment with 0.01% DMSO ± 1.0 μM GLS1i using the improved culture conditions devised here. (a) Live cell numbers as measured by automated microscopy following Hoechst staining and fixation (See Materials and Methods). (b) pH of fresh and spent media samples measured using MColorpHast indicator strips. Concentration of (c) glutamine, (d) glutamate, (e) glucose and (f) lactate in fresh and spent media samples after treatment with or without 1.0 μM GLS1i measured by LC-UV (glutamine and glutamate), Accu-Chek Aviva Blood Glucose Meter System (glucose) and LC-MS (lactate). For each experiment, measurements were performed in triplicate for control and treated conditions. Shown are the mean ± SEM of 3 (A549 cell line; 9 data points per condition) or 2 (H358 cell line; 6 data points per condition) independent experiments. Note that glutamine concentrations in fresh media used for A549 and H358 cells fell by an average of ~27% and ~5% respectively over the duration of the assay. Unadjusted p-values of the differences between control and treated conditions obtained using a two-tailed Student’s t-test are denoted with asterisks: **p ≤ 0.01; ***p ≤ 0.001.
