Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling

Automated single-cell dispensing is incompatible with white adipose tissue (WAT) due to lipid-laden adipocytes. Single-nuclei RNA-Seq permits transcriptional profiling of all cells from WAT. Human WAT faces unique technical challenges in isolating nuclei compared to rodent tissue due to greater extr...

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Detalles Bibliográficos
Autores principales: Whytock, Katie L., Divoux, Adeline, Sun, Yifei, Hopf, Meghan, Yeo, Reichelle X., Pino, Maria F., Yu, GongXin, Smith, Steven R., Walsh, Martin J., Sparks, Lauren M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9876942/
https://www.ncbi.nlm.nih.gov/pubmed/36853719
http://dx.doi.org/10.1016/j.xpro.2023.102054
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author Whytock, Katie L.
Divoux, Adeline
Sun, Yifei
Hopf, Meghan
Yeo, Reichelle X.
Pino, Maria F.
Yu, GongXin
Smith, Steven R.
Walsh, Martin J.
Sparks, Lauren M.
author_facet Whytock, Katie L.
Divoux, Adeline
Sun, Yifei
Hopf, Meghan
Yeo, Reichelle X.
Pino, Maria F.
Yu, GongXin
Smith, Steven R.
Walsh, Martin J.
Sparks, Lauren M.
author_sort Whytock, Katie L.
collection PubMed
description Automated single-cell dispensing is incompatible with white adipose tissue (WAT) due to lipid-laden adipocytes. Single-nuclei RNA-Seq permits transcriptional profiling of all cells from WAT. Human WAT faces unique technical challenges in isolating nuclei compared to rodent tissue due to greater extra-cellular matrix content and larger lipid droplets. In this protocol, we detail how to isolate nuclei from frozen subcutaneous human WAT for single-nuclei RNA-Seq. For complete information on the generation and use of this protocol, please refer to Whytock et al. (2022).(1)
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spelling pubmed-98769422023-01-27 Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling Whytock, Katie L. Divoux, Adeline Sun, Yifei Hopf, Meghan Yeo, Reichelle X. Pino, Maria F. Yu, GongXin Smith, Steven R. Walsh, Martin J. Sparks, Lauren M. STAR Protoc Protocol Automated single-cell dispensing is incompatible with white adipose tissue (WAT) due to lipid-laden adipocytes. Single-nuclei RNA-Seq permits transcriptional profiling of all cells from WAT. Human WAT faces unique technical challenges in isolating nuclei compared to rodent tissue due to greater extra-cellular matrix content and larger lipid droplets. In this protocol, we detail how to isolate nuclei from frozen subcutaneous human WAT for single-nuclei RNA-Seq. For complete information on the generation and use of this protocol, please refer to Whytock et al. (2022).(1) Elsevier 2023-01-20 /pmc/articles/PMC9876942/ /pubmed/36853719 http://dx.doi.org/10.1016/j.xpro.2023.102054 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Whytock, Katie L.
Divoux, Adeline
Sun, Yifei
Hopf, Meghan
Yeo, Reichelle X.
Pino, Maria F.
Yu, GongXin
Smith, Steven R.
Walsh, Martin J.
Sparks, Lauren M.
Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling
title Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling
title_full Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling
title_fullStr Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling
title_full_unstemmed Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling
title_short Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling
title_sort isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9876942/
https://www.ncbi.nlm.nih.gov/pubmed/36853719
http://dx.doi.org/10.1016/j.xpro.2023.102054
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