Oxygen Consumption and Extracellular Acidification Rate in Adipogenic Differentiated Human Mesenchymal Stromal/Stem Cells.

Human mesenchymal stem/stromal cells (hMSCs) constitute a primary cell source in adipogenesis, playing a pivotal role in the generation of new adipocytes. This process is accompanied by substantial metabolic shifts in hMSCs, notably characterized by a transition from glycolytic to oxidative metabolism. Monitoring metabolic alterations during adipogenic differentiation can be achieved through the assessment of extracellular flux parameters, such as the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). In this study, we provide a comprehensive protocol for differentiating hMSCs into the adipogenic lineage, with an emphasis on tracking differentiation stages through extracellular flux measurements. We determined the cellular metabolic status throughout the adipogenic differentiation by assessing OCR and ECAR to gauge mitochondrial respiration and glycolytic activity, respectively, using a Seahorse XFe96 extracellular flux analyzer.