This study explored the impact of different growth medium on the metabolism of human pluripotent stem cells (hPSCs). While these liquid media have been validated for undifferentiated growth of hPSCs, their metabolic impact has not been explored in detail. Using advanced methods of tracking how different nutrients are used by cells, we discovered that these media induce drastic changes in the metabolic state of hPSCs. We observed a major difference in metabolism associated with lipids or fat due to a lack of lipids present in some of the newer, more chemically defined media. Some of these pathways are associated with oxidative stress responses. Supplementation of fat into these media reduced the metabolic activity and increased mitochondrial activity, providing a means for improving the metabolic state of hPSCs and their derivatives.