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

Diferenciação e imagem de adipócitos marrons da fração vascular estromal do tecido adiposo intere...

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

Diferenciação e imagem de adipócitos marrons da fração vascular estromal do tecido adiposo interescapular de camundongos recém-nascidos

Full Text

2,501 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.

Os pré-adipócitos são isolados da fração vascular estromal do tecido adiposo marrom interescapular de camundongos recém-nascidos e diferenciados em células que acumulam gotículas lipídicas, expressam marcadores moleculares e mostram a morfologia mitocondrial de adipócitos marrons maduros. Essas células são posteriormente analisadas por imunofluorescência e microscopia eletrônica de transmissão.

Este protocolo permite a diferenciação eficiente e reprodutível de adipócitos marrons que expressam os principais fatores de transcrição adipogênicos, marcadores de adipócitos maduros e têm potencial termogênico e fenótipo morfológico de adipócitos marrons maduros. Este é um procedimento de diferenciação bifásica simples e replicável para obter células com características de adipócitos marrons maduros a partir de tecido adiposo marrom interescapular de camundongos recém-nascidos. Este protocolo permite estudar os mecanismos de ativação do tecido adiposo marrom como alvo para o tratamento da obesidade.

Tem sido um modelo para determinar os mecanismos envolvidos na fisiopatologia da lipodistrofia. Espera-se que a execução dessa técnica pela primeira vez seja um desafio. É essencial ter um protocolo detalhado e enfatizar pontos críticos para ter resultados bem-sucedidos.

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