Isolation of nuclei from mouse white adipose tissues for single-nucleus genomics

Lipid-filled adipocytes are incompatible with droplet-based single-cell methods, such as 10x Genomics-based technology, thus restricting droplet-based single-cell analyses of adipose tissues to the stromal vascular fraction. To overcome this limitation and obtain cellular and molecular insight into...

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Autores principales: Van Hauwaert, Elvira Laila, Gammelmark, Ellen, Sárvári, Anitta Kinga, Larsen, Lena, Nielsen, Ronni, Madsen, Jesper Grud Skat, Mandrup, Susanne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8220393/
https://www.ncbi.nlm.nih.gov/pubmed/34189477
http://dx.doi.org/10.1016/j.xpro.2021.100612
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author Van Hauwaert, Elvira Laila
Gammelmark, Ellen
Sárvári, Anitta Kinga
Larsen, Lena
Nielsen, Ronni
Madsen, Jesper Grud Skat
Mandrup, Susanne
author_facet Van Hauwaert, Elvira Laila
Gammelmark, Ellen
Sárvári, Anitta Kinga
Larsen, Lena
Nielsen, Ronni
Madsen, Jesper Grud Skat
Mandrup, Susanne
author_sort Van Hauwaert, Elvira Laila
collection PubMed
description Lipid-filled adipocytes are incompatible with droplet-based single-cell methods, such as 10x Genomics-based technology, thus restricting droplet-based single-cell analyses of adipose tissues to the stromal vascular fraction. To overcome this limitation and obtain cellular and molecular insight into adipose tissue composition and plasticity, single-nucleus sequencing-based technologies can be applied. Here, we provide an optimized protocol for nuclei isolation from mouse adipose tissues suitable for single-nucleus RNA sequencing. This allows for transcriptomic profiling of the entire adipose tissue at single-cell resolution. For complete details on the use of this protocol, please refer to Sárvári et al., 2021.
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spelling pubmed-82203932021-06-28 Isolation of nuclei from mouse white adipose tissues for single-nucleus genomics Van Hauwaert, Elvira Laila Gammelmark, Ellen Sárvári, Anitta Kinga Larsen, Lena Nielsen, Ronni Madsen, Jesper Grud Skat Mandrup, Susanne STAR Protoc Protocol Lipid-filled adipocytes are incompatible with droplet-based single-cell methods, such as 10x Genomics-based technology, thus restricting droplet-based single-cell analyses of adipose tissues to the stromal vascular fraction. To overcome this limitation and obtain cellular and molecular insight into adipose tissue composition and plasticity, single-nucleus sequencing-based technologies can be applied. Here, we provide an optimized protocol for nuclei isolation from mouse adipose tissues suitable for single-nucleus RNA sequencing. This allows for transcriptomic profiling of the entire adipose tissue at single-cell resolution. For complete details on the use of this protocol, please refer to Sárvári et al., 2021. Elsevier 2021-06-17 /pmc/articles/PMC8220393/ /pubmed/34189477 http://dx.doi.org/10.1016/j.xpro.2021.100612 Text en © 2021 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
Van Hauwaert, Elvira Laila
Gammelmark, Ellen
Sárvári, Anitta Kinga
Larsen, Lena
Nielsen, Ronni
Madsen, Jesper Grud Skat
Mandrup, Susanne
Isolation of nuclei from mouse white adipose tissues for single-nucleus genomics
title Isolation of nuclei from mouse white adipose tissues for single-nucleus genomics
title_full Isolation of nuclei from mouse white adipose tissues for single-nucleus genomics
title_fullStr Isolation of nuclei from mouse white adipose tissues for single-nucleus genomics
title_full_unstemmed Isolation of nuclei from mouse white adipose tissues for single-nucleus genomics
title_short Isolation of nuclei from mouse white adipose tissues for single-nucleus genomics
title_sort isolation of nuclei from mouse white adipose tissues for single-nucleus genomics
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8220393/
https://www.ncbi.nlm.nih.gov/pubmed/34189477
http://dx.doi.org/10.1016/j.xpro.2021.100612
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