A Protocol to Induce Brown and Beige Adipocyte Differentiation From Murine and Human Adipose-Derived SVF

Adipose cells vary functionally, with white adipocytes storing energy and brown/beige adipocytes generating heat. Mouse and human subcutaneous white adipose tissue (WAT)-derived stromal vascular fraction (SVF) provides mesenchymal stem cells (MSCs) that can be differentiated into thermogenic adipocytes using pharmacological cocktails. After six days of browning induction, these cells exhibited significant upregulation of thermogenic markers (<i>UCP1, Cidea, Dio2, PRDM16</i>) along with adipogenic genes (<i>PPARγ, aP2</i>), showing enhanced thermogenic potential. This in vitro system offers a practical platform to study adipogenesis and thermogenic regulation. Key features • The protocol aims to differentiate murine and human SVF from subcutaneous fat into brown/beige adipocytes for assays. • This approach employs a straightforward methodology that effectively differentiates SVF cells using optimal chemicals, their concentrations, and durations.