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Isolating Brown Adipocytes from Murine Interscapular Brown Adipose Tissue for Gene and Protein Expression Analysis
Published on: March 12, 2021
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Isolation and Characterization of Human Brown Adipocytes
Camilla Scheele1, Tora Ida Henriksen2, Søren Nielsen2
1Novo Nordisk Foundation Center for Basic Metabolic Research, University of Copenhagen, Copenhagen, Denmark. cs@sund.ku.dk.
Methods in Molecular Biology (Clifton, N.J.)
|February 15, 2022
Summary
Researchers developed a method to isolate and differentiate brown adipose tissue (BAT) progenitors from human biopsies. This in vitro model allows for studying thermogenic capacity and molecular mechanisms of BAT.
Area of Science:
- Metabolism and Endocrinology
- Cell Biology
- Tissue Engineering
Background:
- Brown adipose tissue (BAT) plays a crucial role in thermoregulation and energy expenditure.
- Challenges in sampling human BAT in vivo necessitate robust in vitro models.
- Understanding BAT's molecular mechanisms requires accessible cellular models.
Purpose of the Study:
- To establish a protocol for isolating and differentiating human brown adipose tissue (BAT) progenitor cells.
- To characterize the thermogenic capacity of these in vitro differentiated BAT cells.
- To provide a valuable model for mechanistic and molecular studies of human BAT.
Main Methods:
- Isolation of progenitor cells from the stromal vascular fraction of human BAT biopsies.
- In vitro differentiation of isolated progenitor cells into functional brown adipocytes.
- Characterization of thermogenic capacity using qPCR for gene expression, Seahorse XFe96 Analyzer for mitochondrial uncoupling, and RNAscope® Technology for UCP1 expression.
Main Results:
- Successful isolation and differentiation of human BAT progenitor cells in vitro.
- Demonstrated norepinephrine-induced thermogenic gene expression and mitochondrial uncoupling.
- Confirmed UCP1 expression, a key marker of brown adipocyte thermogenic function.
Conclusions:
- The described protocol provides a reliable method for generating an in vitro model of human brown adipose tissue.
- This model enables detailed mechanistic and molecular investigations into BAT function and thermogenesis.
- The characterized cells offer a valuable resource for research into obesity, metabolic diseases, and energy balance.
