Protocol for measuring mitochondrial respiration in mature human adipocytes using the Seahorse XF analyzer

Primary mature human adipocytes are an important tool bridging basic research and clinical medicine by reflecting human biological diversity. Here, we present a protocol for measuring the oxygen consumption rate of mature adipocytes. We describe the steps for isolation of adipocytes from adipose tissue samples, Matrigel embedding, and Seahorse analysis, as well as normalization and data analysis. This protocol enables the investigation of the metabolic function of primary mature human adipocytes in response to drugs or genetic modifications.