Differentiation of human subcutaneous adipocytes and measurement of lipolytic function induced by GIP or LY3437943

Human subcutaneous adipocytes are an attractive therapeutic target in regulating overall physiological homeostasis. However, the differentiation of primary human adipose-derived models remains challenging. Here, we present a protocol to differentiate primary subcutaneous adipose-derived preadipocytes from human subcutaneous adipocytes and to measure lipolytic activity. We describe steps for seeding of subcutaneous preadipocytes, removal of growth factors, induction and maturation of adipocytes, removal of serum/phenol red in media, and treatment of mature adipocytes. We then detail glycerol measurement in conditioned media and its interpolation. For complete details on the use and execution of this protocol, please refer to Coskun et al.(1)