JoVE Journal > Biology
Summary
Abstract
Introduction
Protocol
Results
Discussion
Disclosures
Acknowledgements
Materials
References
Biology

Mensuração da Taxa de Lipólise em Tecido Adiposo Murino Ex Vivo e Pré-adipócitos Primários Diferenciados In Vitro

Published: March 17, 2023
doi:
10.3791/65106
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,
1Weill Center for Metabolic Health, Department of Medicine,Weill Cornell Medicine

Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

Disclosures

The authors have nothing to disclose.

Acknowledgements

Materials

24-Well tissue culture treated plateCorning Inc3527Must be tissue culture treated for adipocyte differntiation
48-Well flat bottom plate with lidCorning Inc353078Can be tissue culture treated
6-Well flat bottom plate with lidCorning Inc353046Can be tissue culture treated
96-Well PCR PlateUSA sceintific1402-9100Any conical 0.2 mL PCR plate will be convenient 
Bovine Serum AlbuminSigma AldrichA9418FFA free BSA such as A8806, is also commonly used. The BSA should not have detectable FFA, also lot to lot variations in BSA can impact the observed rate of lipolysis
CL-316,243Sigma AldrichC5976CAS #: 138908-40-4 availaible from other suppliers
CO2 incubatorPHCBIMCO-170AICUVHCO2 should ideally be set to 10% for adipose tissue, however 5% CO2 will also work
DMEM, low glucose, no phenol redThermofischer11054020Any phenol red free media should work, DMEM/F12, RPMI, but should contain volatile buffering capacity, i.e. biocarbonate
FFA-free Bovine serum albuminEquitech-Bio, Inc, BAH66
Free Glycerol ReagentSigma AldrichF6428
Glycerol Standard SolutionSigma AldrichG7793 This can also be made by diluting glycerol to the desired concentration
HR Series NEFA Standard SolutionFujifilm276-76491
HR Series NEFA-HR (2) Color Reagent AFujifilm999-34691
HR Series NEFA-HR (2) Color Reagent BFujifilm991-34891
HR Series NEFA-HR (2) Solvent A Fujifilm995-34791
HR Series NEFA-HR (2) Solvent B Fujifilm993-35191
Microbiological IncubatorFischer ScientificS28668Any incubator at 37C can be used
Nunc MicroWell 96-Well PlatesThermo Scientific269620Any optically clear, flat bottom 96-well plate works
Silicone Laboratory Benchtop MatVWR76045-300Glass plate can also be used. Absorbant surfaces are not recommended
Spectrophotometer/Microplate ReaderMolecular devicesSpectraMax i3x Any plate reader that can read at 540, 550 and 660 mm will work
V Bovine serum albuminSigma-Aldrich810531
WypAll X70 WipersKimberly-Clark41200Any high quality paper towel will work
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References

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LipolysisPrimary PreadipocytesAdipocyte DifferentiationBovine Serum AlbuminBSA MediumDulbecco’s Modified Eagle’s MediumDMEMInsulin TreatmentSerial SamplingMeasurement ErrorAdipogenic Cell LinesControl MediaStimulation Media
Measuring the Rate of Lipolysis in Ex Vivo Murine Adipose Tissue and Primary Preadipocytes Differentiated In Vitro

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Bridge-Comer, P. E., Reilly, S. M. Measuring the Rate of Lipolysis in Ex Vivo Murine Adipose Tissue and Primary Preadipocytes Differentiated In Vitro. J. Vis. Exp. (193), e65106, doi:10.3791/65106 (2023).
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