Differentiation and Imaging of Brown Adipocytes from the Stromal Vascular Fraction of Interscapular Adipose Tissue from Newborn Mice

Ana-Mar&#237;a Figueroa; Francisca Stolzenbach; Pablo Tapia; V&#237;ctor Cort&#233;s

doi:10.3791/64604

Differenziazione e imaging di adipociti marroni dalla frazione vascolare stromale del tessuto adi...

JoVE Journal
Biology
Author Produced

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Biology

Differentiation and Imaging of Brown Adipocytes from the Stromal Vascular Fraction of Interscapular Adipose Tissue from Newborn Mice

Differenziazione e imaging di adipociti marroni dalla frazione vascolare stromale del tessuto adiposo interscapolare da topi neonati

Full Text

2,543 Views

04:46 min

February 3, 2023

DOI: 10.3791/64604-v

Ana-María Figueroa¹, Francisca Stolzenbach¹, Pablo Tapia¹, Víctor Cortés¹

¹Department of Nutrition, Diabetes and Metabolism,Pontificia Universidad Católica de Chile

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study provides a detailed protocol for the differentiation of preadipocytes into mature brown adipocytes using interscapular brown adipose tissue harvested from newborn mice. The differentiated cells display characteristic lipid accumulation, express key adipogenic markers, and exhibit mitochondrial adaptations indicative of thermogenic functionality.

Key Study Components

Research Area

Adipocyte biology
Obesity research
Mechanisms of thermogenesis

Background

Investigation of brown adipose tissue's role in energy metabolism
Modeling conditions related to obesity and lipodystrophy
Importance of preadipocyte differentiation for studying adipogenesis

Methods Used

Immunofluorescence and transmission electron microscopy
Interscapular brown adipose tissue from newborn mice
Cell differentiation protocols and assays for mitochondrial characteristics

Main Results

Successful differentiation of preadipocytes into mature adipocytes marked by lipid droplet accumulation
Enhanced expression of adipogenic transcription factors and thermogenic markers on differentiation day seven
Characterization of mitochondrial morphology changes associated with adipogenesis

Conclusions

This protocol enables the reproducible study of brown adipocyte differentiation and functions.
Relevance to understanding obesity and potential therapeutic targets.

Frequently Asked Questions

What are preadipocytes?

Preadipocytes are undifferentiated cells that can develop into adipocytes, which store fat.

Why focus on brown adipocytes?

Brown adipocytes play a key role in thermogenesis and energy expenditure, making them important in obesity research.

What techniques are used to analyze differentiated cells?

Immunofluorescence and transmission electron microscopy are employed for detailed cellular analysis.

What markers indicate successful differentiation?

Key markers include PPAR-gamma, C/EBP-alpha, and UCP1, which are significantly upregulated in mature brown adipocytes.

What is the importance of mitochondrial morphology in this study?

Mitochondrial morphology is crucial for assessing the thermogenic potential and functionality of differentiated brown adipocytes.

How can this research be applied?

It can help in understanding obesity and developing potential therapeutic strategies for metabolic disorders.

I preadipociti sono isolati dalla frazione vascolare stromale del tessuto adiposo bruno interscapolare di topi neonati e differenziati in cellule che accumulano goccioline lipidiche, esprimono marcatori molecolari e mostrano la morfologia mitocondriale degli adipociti bruni maturi. Queste cellule vengono ulteriormente analizzate mediante immunofluorescenza e microscopia elettronica a trasmissione.

Questo protocollo consente la differenziazione efficiente e riproducibile di adipociti bruni che esprimono fattori chiave di trascrizione adipogenica, marcatori adipocitari maturi e hanno un potenziale termogenico e un fenotipo morfologico degli adipociti bruni maturi. Si tratta di una procedura di differenziazione in due fasi semplice e replicabile per ottenere cellule con caratteristiche di adipociti bruni maturi da tessuto adiposo bruno interscapolare di topi neonati. Questo protocollo permette di studiare i meccanismi di attivazione del tessuto adiposo bruno come bersaglio per il trattamento dell'obesità.

È stato un modello per determinare i meccanismi coinvolti nella fisiopatologia della lipodistrofia. L'esecuzione di questa tecnica per la prima volta dovrebbe essere impegnativa. È essenziale avere un protocollo dettagliato e sottolineare i punti critici per avere risultati di successo.

View the full transcript and gain access to thousands of scientific videos