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

Ajit Regmi; William Roell

doi:10.1016/j.xpro.2023.102304

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

STAR Protoc. 2023 May 12;4(2):102304. doi: 10.1016/j.xpro.2023.102304. Online ahead of print.

Authors

Ajit Regmi¹, William Roell²

Affiliations

¹ Insulin Discovery, Eli Lilly and Company, Indianapolis, IN 46225, USA. Electronic address: regmi_ajit@lilly.com.
² Insulin Discovery, Eli Lilly and Company, Indianapolis, IN 46225, USA. Electronic address: roell_william_c@lilly.com.

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.¹.

Keywords: Cell Biology; Metabolism.