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J Vis Exp. 2013 Mar 28;(73). doi: 10.3791/50191.

Isolation and differentiation of stromal vascular cells to beige/brite cells.

Author information

1
UCSF Diabetes Center and Department of Cell and Tissue Biology, University of California, San Francisco, CA, USA.

Abstract

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 PPARĪ³ 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.

PMID:
23568137
PMCID:
PMC3641667
DOI:
10.3791/50191
[Indexed for MEDLINE]
Free PMC Article

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