A Method to Induce Brown/Beige Adipocyte Differentiation from Murine Preadipocytes

Author(s)

Rocha, Andrea Livia ^{[1, 2, 3, 4]} ; Guerra, Beatriz Alves ^[3] ; Boucher, Jeremie ^{[5, 6, 7]} ; Mori, Marcelo A. ^{[1, 2, 3, 4, 8, 9]}

Affiliation

^[1] Univ Estadual Campinas, Inst Biol, Dept Biochem & Tissue Biol, Campinas - Brazil

^[2] Univ Estadual Campinas, Inst Biol, Program Genet & Mol Biol, Campinas - Brazil

^[3] Univ Fed Sao Paulo, Sao Paulo Sch Med, Dept Biophys, Sao Paulo - Brazil

^[4] Univ Fed Sao Paulo, Program Biotechnol, Sao Paulo - Brazil

^[5] Univ Gothenburg, Wallenberg Ctr Mol & Translat Med, Gothenburg - Sweden

^[6] Univ Gothenburg, Lundberg Lab Diabet Res, Gothenburg - Sweden

^[7] Evotec Int GmbH, Metab Dis, Gottingen - Germany

^[8] Univ Estadual Campinas, Obes & Comorbid Res Ctr OCRC, Campinas - Brazil

^[9] Expt Med Res Cluster EMRC, Campinas - Brazil

• Document type: Journal article
• Source: BIO-PROTOCOL; v. 11, n. 24 DEC 20 2021.
• Web of Science Citations: 0

Abstract

Adipocytes exhibit different morphological and functional characteristics, depending on their anatomical location, developmental origin, and stimulus. While white adipocytes tend to accumulate energy as triglycerides, brown and beige adipocytes tend to direct carbon sources to fuel thermogenesis. White and beige adipocytes originate from common progenitor cells, which are distinct from brown adipocyte precursors. Having a method to study white vs. beige vs. brown adipocyte differentiation may help to unveil the mechanisms driving distinct adipogenic programs. Preadipocytes can be cultured and differentiated in vitro using a combination of compounds to stimulate adipogenesis. Here, we describe and compare protocols designed to stimulate adipocyte differentiation and induce brown/beige-like or white-like characteristics in differentiating adipocytes. The protocols consist in exposing murine preadipocytes to pharmacological stimuli aimed at triggering adipogenesis and inducing (or not) a thermogenic gene expression program. After 8 days of differentiation with a pro-browning cocktail, immortalized preadipocytes isolated from interscapular brown fat (9B cells) or inguinal white fat (9W cells) from the same mouse expressed higher levels of brown/beige adipocyte markers (e.g., Ucp1) and pan-adipocyte differentiation markers (e.g., Pparg, Cebpa and aP2) when compared to the same cells differentiated with a cocktail that lacked brown/beige adipogenic inducers (i.e., rosiglitazone, T3, and indomethacin). Consistent with a higher thermogenic potential of brown vs. beige adipocytes, differentiated 9B cells expressed higher Ucp1 levels than differentiated 9W cells. This simple protocol may help researchers to understand mechanisms of adipogenesis and how adipocytes become thermogenic. (AU)

• FAPESP's process: 15/03292-8 - Study of the role of GCN1 in the genesis of metabolic diseases in mice
• Grantee: Beatriz Alves Guerra
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 16/12294-7 - Dicer signalling in adipocytes controlling immune function and activation of beige cells in adipose tissue
• Grantee: Andréa Livia Silva Rocha
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 17/01184-9 - CAMeLEOm: cross-species analysis of metabolic, lifespan effects and omics of dietary restriction mimetics
• Grantee: Marcelo Alves da Silva Mori
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 12/07259-7 - Evaluation of miRNA Processing in Adipose Tissue of Mice Subjected to Caloric Restriction
• Grantee: Beatriz Alves Guerra
• Support Opportunities: Scholarships in Brazil - Master