Isolation of Immune Cells from Adipose Tissue for Flow Cytometry
Isolation of Immune Cells from Adipose Tissue for Flow Cytometry
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DOI：
10.1007/978-1-4939-6820-6_6
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发表时间：
2017-01-01
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影响因子：
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通讯作者：
Brestoff, Jonathan R.
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作者：
Brestoff, Jonathan R.
米色和棕色脂肪细胞分别是维持白色脂肪组织（WAT）和棕色脂肪组织（BAT）内代谢稳态所必需的产热细胞。新兴的研究表明，各种免疫细胞类型，如交替激活的巨噬细胞（AAMacs），嗜酸性粒细胞和第2组先天性淋巴样细胞（ILC 2）关键调节米色和/或棕色脂肪细胞的发育和激活，以防止肥胖和维持核心体温。这些发现表明，米色和棕色脂肪组织的研究可能受益于传统的免疫学方法，如WAT和BAT内免疫细胞的流式细胞术。本文的目的是描述一种有效的方法来分离免疫细胞从大量的脂肪组织样本平行。然后可以通过多种方法评估用该方案分离的细胞的组成、表型和活化状态，包括但不限于流式细胞术。作为一个例子，这篇文章简要介绍了一种方法，以确定AAMacs，嗜酸性粒细胞和ILC 2的脂肪组织。
Beige and brown adipocytes are thermogenic cells essential for maintaining metabolic homeostasis within white adipose tissue (WAT) and brown adipose tissue (BAT), respectively. Emerging studies indicate that various immune cell types such as alternatively activated macrophages (AAMacs), eosinophils, and group 2 innate lymphoid cells (ILC2s) critically regulate beige and/or brown adipocyte development and activation to protect against obesity and maintain core body temperature. These findings suggest that studies of beige and brown adipose tissue may benefit from traditional immunologic approaches such as flow cytometry of immune cells residing within WAT and BAT. The purpose of this article is to describe an efficient method to isolate immune cells from numerous adipose tissue samples in parallel. The composition, phenotype, and activation state of cells isolated with this protocol may then be assessed by multiple methods including but not limited to flow cytometry. As an example, this article briefly describes a method to identify AAMacs, eosinophils, and ILC2s in adipose tissues.