Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling
Summary: 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 gre...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723000126 |
| _version_ | 1797946162665226240 |
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| author | Katie L. Whytock Adeline Divoux Yifei Sun Meghan Hopf Reichelle X. Yeo Maria F. Pino GongXin Yu Steven R. Smith Martin J. Walsh Lauren M. Sparks |
| author_facet | Katie L. Whytock Adeline Divoux Yifei Sun Meghan Hopf Reichelle X. Yeo Maria F. Pino GongXin Yu Steven R. Smith Martin J. Walsh Lauren M. Sparks |
| author_sort | Katie L. Whytock |
| collection | DOAJ |
| description | Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-10T21:06:31Z |
| format | Article |
| id | doaj.art-6f05c012201949db8ce2f85ec5f3f189 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-10T21:06:31Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-6f05c012201949db8ce2f85ec5f3f1892023-01-22T04:42:07ZengElsevierSTAR Protocols2666-16672023-03-0141102054Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profilingKatie L. Whytock0Adeline Divoux1Yifei Sun2Meghan Hopf3Reichelle X. Yeo4Maria F. Pino5GongXin Yu6Steven R. Smith7Martin J. Walsh8Lauren M. Sparks9Translational Research Institute, AdventHealth, Orlando, FL 32804, USA; Corresponding authorTranslational Research Institute, AdventHealth, Orlando, FL 32804, USAIcahn School of Medicine at Mount Sinai, New York, NY 10029, USATranslational Research Institute, AdventHealth, Orlando, FL 32804, USATranslational Research Institute, AdventHealth, Orlando, FL 32804, USATranslational Research Institute, AdventHealth, Orlando, FL 32804, USATranslational Research Institute, AdventHealth, Orlando, FL 32804, USATranslational Research Institute, AdventHealth, Orlando, FL 32804, USAIcahn School of Medicine at Mount Sinai, New York, NY 10029, USATranslational Research Institute, AdventHealth, Orlando, FL 32804, USA; Corresponding authorSummary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723000126Cell isolationSingle CellRNAseqMetabolism |
| spellingShingle | Katie L. Whytock Adeline Divoux Yifei Sun Meghan Hopf Reichelle X. Yeo Maria F. Pino GongXin Yu Steven R. Smith Martin J. Walsh Lauren M. Sparks Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling STAR Protocols Cell isolation Single Cell RNAseq Metabolism |
| 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 | Cell isolation Single Cell RNAseq Metabolism |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723000126 |
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