Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 73, Pages -Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/50191
Keywords
Cellular Biology; Issue 73; Medicine; Anatomy; Physiology; Molecular Biology; Surgery; Adipose Tissue; Adipocytes; Transcription Factors; Cell Differentiation; Obesity; Diabetes; brown adipose tissue; beige/brite cells; primary adipocytes; stromal-vascular fraction; differentiation; uncoupling protein 1; rosiglitazone; differentiation; cells; isolation; fat; animal model
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Funding
- NIH [DK087853]
- Program for Breakthrough Biomedical Research
- Asubio Pharm Inc.
- SHARE PhD fellowships from The University of Copenhagen
- EU FP7 project DIABAT [HEALTH-F2-2011-278373]
- DERC center grant [NIH P30 DK063720]
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Brown adipocytes have the ability to uncouple the respiratory chain in mitochondria and dissipate chemical energy as heat. Development of UCP1-positive brown adipocytes in white adipose tissues (so called beige or brite cells) is highly induced by a variety of environmental cues such as chronic cold exposure or by PPAR. agonists, therefore, this cell type has potential as a therapeutic target for obesity treatment. Although most immortalized adipocyte lines cannot recapitulate the process of browning of white fat in culture, primary adipocytes isolated from stromal vascular fraction in subcutaneous white adipose tissue (WAT) provide a reliable cellular system to study the molecular control of beige/brite cell development. Here we describe a protocol for effective isolation of primary preadipocytes and for inducing differentiation to beige/brite cells in culture. The browning effect can be assessed by the expression of brown fat-selective markers such as UCP1.
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