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Mesure du taux de lipolyse dans le tissu adipeux murin ex vivo et les préadipocytes prim...
Mesure du taux de lipolyse dans le tissu adipeux murin ex vivo et les préadipocytes prim...
JoVE Journal
Biology
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JoVE Journal Biology
Measuring the Rate of Lipolysis in Ex Vivo Murine Adipose Tissue and Primary Preadipocytes Differentiated In Vitro

Mesure du taux de lipolyse dans le tissu adipeux murin ex vivo et les préadipocytes primaires différenciés in vitro

Full Text
3,864 Views
09:41 min
March 17, 2023

DOI: 10.3791/65106-v

Pania E. Bridge-Comer1, Shannon M. Reilly1

1Weill Center for Metabolic Health, Department of Medicine,Weill Cornell Medicine

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

Overview

This study presents a protocol for measuring the lipolytic rates in adipocytes, utilizing both cultured cells and ex vivo adipose tissue from murine models. The findings significantly demonstrate the differences in lipolytic rates under basal and stimulated conditions.

Key Study Components

Research Area

  • Adipocyte metabolism
  • Lipolysis measurement
  • Cutting-edge metabolic assays

Background

  • Understanding triglyceride lipolysis is crucial for insights into metabolic processes.
  • Utilizing murine models aids in the exploration of adipose tissue responses.
  • The relevance of free fatty acids and glycerol as metabolic byproducts is emphasized.

Methods Used

  • Serial sampling and media collection from cultured adipocyte and ex vivo tissue.
  • Murine adipose tissue was used for comparative analysis.
  • Enzymatic assays for quantifying free fatty acid and glycerol production.

Main Results

  • Stimulated lipolytic rates were significantly higher compared to basal rates.
  • FFA-to-glycerol molar ratios indicated varying sources of glycerol amidst different conditions.
  • Results validate the protocol's applicability across model systems.

Conclusions

  • The study successfully illustrates how lipolytic rates can be precisely measured in both in vitro and ex vivo environments.
  • These findings have broad implications for understanding metabolic disorders associated with adipose tissue function.

Frequently Asked Questions

What is the main focus of the study?
The study focuses on the protocol for measuring lipolysis in adipocytes from both cultured and ex vivo tissues.
Why is triglyceride lipolysis important?
Triglyceride lipolysis is essential for understanding energy metabolism and its dysregulation in metabolic diseases.
What organisms are used in the study?
Murine models are used for the comparative analysis of adipose tissue.
How are lipolytic rates measured?
Lipolytic rates are measured using serial sampling and enzymatic assays to quantify free fatty acids and glycerol.
What were the key findings regarding FFA and glycerol ratios?
In stimulated conditions, the FFA-to-glycerol ratio was about three, indicating effective lipolysis without significant reuptake.
How might this research contribute to metabolic studies?
This research provides a validated approach to measure lipolytic activity, which can be crucial for studying metabolic diseases and treatments.

La lipolyse des triglycérides dans les adipocytes est un processus métabolique important entraînant la libération d’acides gras libres et de glycérol. Ici, nous fournissons un protocole détaillé pour mesurer la lipolyse basale et stimulée dans les adipocytes et le tissu adipeux ex vivo de souris.

Ce protocole détaille la détermination du taux de lipolyse adipocytaire dans les adipocytes en culture ou le tissu adipeux ex vivo, permettant de comparer les taux lipolytiques entre modèles murins ou traitements. Ce protocole utilise l’échantillonnage en série, ce qui permet de valider en interne que la lipolyse est mesurée dans la phase linéaire et réduit l’erreur de mesure. Avec optimisation, ce protocole peut être utilisé pour mesurer la lipolyse dans le tissu adipeux brun, le tissu adipeux d’autres organismes ou les lignées cellulaires adipogènes.

Les tissus ex vivo doivent être coupés en petits morceaux de taille et de forme constantes pour permettre la séquestration des acides gras libres libérés par le BSA dans le milieu. Pour commencer, préparez le milieu BSA à 5% en dissolvant cinq grammes d’albumine sérique bovine, ou BSA, dans 100 millilitres de milieu d’aigle modifié de Dulbecco, ou DMEM, sans rouge de phénol. Remuer doucement la solution pour dissoudre le BSA.

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