Optimization of methods and treatment conditions for studying effects of fatty acids on cell growth

Antiproliferative properties of molecular regulators of lipid metabolism ha ve been increasingly studied during recent years. Discussion is ongoing con cerning optimal treatment conditions and assays used for monitoring prolife ration and cytotoxicity. The objective of the present work was to optimize methods and treatment conditions used for studying antiproliferative effect s of fatty acids and analogs, represented by palmitic acid (PA) and the bet a -oxidation-restricred fatty acid analog tetradecylthioacetic acid (TTA), in rat (BT4Cn) and human (D54Mg and GaMg) glioma cell lines. Changes in [H- 3]thymidine incorporation preceded changes in cell number in TTA-treated gl ioma cell cultures, and the growth inhibition was more significantly expres sed by [H-3]thymidine incorporation than cell number. Addition of bovine se rum albumin decreased cellular fatty acid uptake and reduced the effects of TTA and PA on [H-3]thymidine incorporation. Determination of the antiproli ferative effect of TTA in BT4Cn cells by MTT conversion and [H-3]thymidine incorporation yielded concordant results. TTA-mediated reduction in cell nu mber corresponded to reduction in cellular protein and total DNA content in BT4Cn cells. Reduced growth potential in TTA-treated multicellular D54Mg a nd GaMg spheroids supported the findings from monolayer cultures. In conclu sion, cell density, treatment period, fatty acid administration, and method s for growth determination may profoundly influence the outcome of cell gro wth experiments. Thus, experimental conditions should be carefully controll ed when performing cell growth experiments, and effects on cell growth shou ld preferably be confirmed by different methods.