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

STAR protocols2023PMID: 37178114

View on PubMed DOI: 10.1016/j.xpro.2023.102304

Plain Language Summary

Describes a laboratory protocol for differentiating primary human subcutaneous preadipocytes into mature adipocytes and measuring lipolytic function. Uses LY3437943 (retatrutide) and GIP to test adipocyte biology, establishing that retatrutide activates lipolysis in human fat cells—a mechanism contributing to its superior fat loss.

Abstract

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..

Authors

Regmi, Ajit; Roell, William

Keywords

Cell BiologyMetabolism