Preparation of adipose progenitor cells from mouse epididymal adipose tissues

Dong Seong Cho; Jason D. Doles

doi:10.3791/61694

Preparation of adipose progenitor cells from mouse epididymal adipose tissues

Dong Seong Cho, Jason D. Doles

Biochemistry and Molecular Biology

Research output: Contribution to journal › Article › peer-review

Abstract

Obesity and metabolic disorders such as diabetes, heart disease, and cancer, are all associated with dramatic adipose tissue remodeling. Tissue-resident adipose progenitor cells (APCs) play a key role in adipose tissue homeostasis and can contribute to the tissue pathology. The growing use of single cell analysis technologies – including single-cell RNA-sequencing and single-cell proteomics – is transforming the stem/progenitor cell field by permitting unprecedented resolution of individual cell expression changes within the context of population-or tissue-wide changes. In this article, we provide detailed protocols to dissect mouse epididymal adipose tissue, isolate single adipose tissue-derived cells, and perform fluorescence activated cell sorting (FACS) to enrich for viable Sca1⁺ /CD31^- /CD45^- /Ter119^- APCs. These protocols will allow investigators to prepare high quality APCs suitable for downstream analyses such as single cell RNA sequencing.

Original language	English (US)
Article number	e61694
Pages (from-to)	1-7
Number of pages	7
Journal	Journal of Visualized Experiments
Volume	2020
Issue number	162
DOIs	https://doi.org/10.3791/61694
State	Published - Aug 2020

ASJC Scopus subject areas

Neuroscience(all)
Chemical Engineering(all)
Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)

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title = "Preparation of adipose progenitor cells from mouse epididymal adipose tissues",

abstract = "Obesity and metabolic disorders such as diabetes, heart disease, and cancer, are all associated with dramatic adipose tissue remodeling. Tissue-resident adipose progenitor cells (APCs) play a key role in adipose tissue homeostasis and can contribute to the tissue pathology. The growing use of single cell analysis technologies – including single-cell RNA-sequencing and single-cell proteomics – is transforming the stem/progenitor cell field by permitting unprecedented resolution of individual cell expression changes within the context of population-or tissue-wide changes. In this article, we provide detailed protocols to dissect mouse epididymal adipose tissue, isolate single adipose tissue-derived cells, and perform fluorescence activated cell sorting (FACS) to enrich for viable Sca1+ /CD31- /CD45- /Ter119- APCs. These protocols will allow investigators to prepare high quality APCs suitable for downstream analyses such as single cell RNA sequencing.",

author = "Cho, {Dong Seong} and Doles, {Jason D.}",

note = "Funding Information: We acknowledge the Mayo Clinic Microscopy Cell Analysis Core Flow Cytometry Facility for assistance with FACS sorting. Publisher Copyright: {\textcopyright} 2020 JoVE Journal of Visualized Experiments.",

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AU - Cho, Dong Seong

AU - Doles, Jason D.

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N2 - Obesity and metabolic disorders such as diabetes, heart disease, and cancer, are all associated with dramatic adipose tissue remodeling. Tissue-resident adipose progenitor cells (APCs) play a key role in adipose tissue homeostasis and can contribute to the tissue pathology. The growing use of single cell analysis technologies – including single-cell RNA-sequencing and single-cell proteomics – is transforming the stem/progenitor cell field by permitting unprecedented resolution of individual cell expression changes within the context of population-or tissue-wide changes. In this article, we provide detailed protocols to dissect mouse epididymal adipose tissue, isolate single adipose tissue-derived cells, and perform fluorescence activated cell sorting (FACS) to enrich for viable Sca1+ /CD31- /CD45- /Ter119- APCs. These protocols will allow investigators to prepare high quality APCs suitable for downstream analyses such as single cell RNA sequencing.

AB - Obesity and metabolic disorders such as diabetes, heart disease, and cancer, are all associated with dramatic adipose tissue remodeling. Tissue-resident adipose progenitor cells (APCs) play a key role in adipose tissue homeostasis and can contribute to the tissue pathology. The growing use of single cell analysis technologies – including single-cell RNA-sequencing and single-cell proteomics – is transforming the stem/progenitor cell field by permitting unprecedented resolution of individual cell expression changes within the context of population-or tissue-wide changes. In this article, we provide detailed protocols to dissect mouse epididymal adipose tissue, isolate single adipose tissue-derived cells, and perform fluorescence activated cell sorting (FACS) to enrich for viable Sca1+ /CD31- /CD45- /Ter119- APCs. These protocols will allow investigators to prepare high quality APCs suitable for downstream analyses such as single cell RNA sequencing.

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Preparation of adipose progenitor cells from mouse epididymal adipose tissues

Abstract

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