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

Elvira Laila Van Hauwaert; Ellen Gammelmark; Anitta Kinga Sárvári; Lena Larsen; Ronni Nielsen; Jesper Grud Skat Madsen; Susanne Mandrup

doi:10.1016/j.xpro.2021.100612

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

Elvira Laila Van Hauwaert^*, Ellen Gammelmark, Anitta Kinga Sárvári, Lena Larsen, Ronni Nielsen, Jesper Grud Skat Madsen, Susanne Mandrup^*

^*Corresponding author for this work

University of Southern Denmark

Research output: Contribution to journal › Journal article › Research › peer-review

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Abstract

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.

Original language	English
Article number	100612
Journal	STAR Protocols
Volume	2
Issue number	3
Number of pages	12
ISSN	2666-1667
DOIs	https://doi.org/10.1016/j.xpro.2021.100612
Publication status	Published - 17. Sept 2021

Bibliographical note

Publisher Copyright:
© 2021 The Author(s)

Keywords

Cell isolation
Gene Expression
Genomics
Metabolism
RNA-seq
Single Cell

Documents & Links

10.1016/j.xpro.2021.100612Licence: CC BY-NC-ND

Open Access VersionFinal published version, 2.61 MBLicence: CC BY-NC-ND

1 Ph.D. thesis

Adipose tissue plasticity in obesity
Hauwaert, E. L. V., 10. Feb 2022, Syddansk Universitet. Det Naturvidenskabelige Fakultet. 99 p.
Research output: Thesis › Ph.D. thesis

Open Access
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Cite this

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title = "Isolation of nuclei from mouse white adipose tissues for single-nucleus genomics",

abstract = "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 S{\'a}rv{\'a}ri et al., 2021.",

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T1 - Isolation of nuclei from mouse white adipose tissues for single-nucleus genomics

AU - Van Hauwaert, Elvira Laila

AU - Gammelmark, Ellen

AU - Sárvári, Anitta Kinga

AU - Larsen, Lena

AU - Nielsen, Ronni

AU - Madsen, Jesper Grud Skat

AU - Mandrup, Susanne

PY - 2021/9/17

Y1 - 2021/9/17

N2 - 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.

AB - 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.

KW - Cell isolation

KW - Gene Expression

KW - Genomics

KW - Metabolism

KW - RNA-seq

KW - Single Cell

U2 - 10.1016/j.xpro.2021.100612

DO - 10.1016/j.xpro.2021.100612

M3 - Journal article

C2 - 34189477

AN - SCOPUS:85108000529

SN - 2666-1667

VL - 2

JO - STAR Protocols

JF - STAR Protocols

IS - 3

M1 - 100612

ER -

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

Abstract

Bibliographical note

Keywords

Documents & Links

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Related research output

Adipose tissue plasticity in obesity

Cite this