NDLI: Isolation of adipose tissue immune cells.

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Isolation of adipose tissue immune cells.

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Year: 2013 Issue: 74

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Isolation of adipose tissue immune cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Orr, Jeb S. Kennedy, Arion J. Hasty, Alyssa H.
Description	Country affiliation: United States Author Affiliation: Orr JS ( Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, USA. alyssa.hasty@vanderbilt.edu)
Abstract	The discovery of increased macrophage infiltration in the adipose tissue (AT) of obese rodents and humans has led to an intensification of interest in immune cell contribution to local and systemic insulin resistance. Isolation and quantification of different immune cell populations in lean and obese AT is now a commonly utilized technique in immunometabolism laboratories; yet extreme care must be taken both in stromal vascular cell isolation and in the flow cytometry analysis so that the data obtained is reliable and interpretable. In this video we demonstrate how to mince, digest, and isolate the immune cell-enriched stromal vascular fraction. Subsequently, we show how to antibody label macrophages and T lymphocytes and how to properly gate on them in flow cytometry experiments. Representative flow cytometry plots from low fat-fed lean and high fat-fed obese mice are provided. A critical element of this analysis is the use of antibodies that do not fluoresce in channels where AT macrophages are naturally autofluorescent, as well as the use of proper compensation controls.
File Format	HTM / HTML
e-ISSN	1940087X
DOI	10.3791/50707
Journal	Journal of Visualized Experiments
Issue Number	75
Language	English
Publisher	MyJove Corp.
Publisher Date	2013-05-22
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Adipose Tissue Cytology Immunology Flow Cytometry Macrophages T-lymphocytes Animals Mice Mice, Inbred C57bl Obesity Pathology Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Video-audio Media
Content Type	Text
Resource Type	Article

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