Isolation and Differentiation of Stromal Vascular Cells to Beige/Brite Cells
Isolation and Differentiation of Stromal Vascular Cells to Beige/Brite Cells
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DOI：
10.3791/50191
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发表时间：
2013-03-01
影响因子：
1.2
通讯作者：
Kajimura, Shingo
中科院分区：
文献类型：
--
作者：
Aune, Ulrike Liisberg;Ruiz, Lauren;Kajimura, Shingo
棕色脂肪细胞具有解偶联线粒体中的呼吸链并将化学能作为热量耗散的能力。白色脂肪组织中UCP 1阳性棕色脂肪细胞（所谓的米色或棕色细胞）的发育受到多种环境因素如慢性冷暴露或PPAR的高度诱导。因此，这种细胞类型具有作为肥胖症治疗的治疗靶点的潜力。虽然大多数永生化脂肪细胞系不能重现培养中白色脂肪的“布朗宁”过程，但从皮下白色脂肪组织（WAT）中的基质血管部分分离的原代脂肪细胞为研究米色/棕色细胞发育的分子控制提供了可靠的细胞系统。在这里，我们描述了一个协议，有效地分离原代前脂肪细胞和诱导分化为米色/brite细胞培养。布朗宁效应可以通过褐脂选择性标记物如UCP 1的表达来评估。
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.