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^*

^*Kontaktforfatter

Syddansk Universitet

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › 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.

Originalsprog	Engelsk
Artikelnummer	100612
Tidsskrift	STAR Protocols
Vol/bind	2
Udgave nummer	3
Antal sider	12
ISSN	2666-1667
DOI	https://doi.org/10.1016/j.xpro.2021.100612
Status	Udgivet - 17. sep. 2021

Bibliografisk note

Funding Information:
The work was supported by grants from the Danish National Research Foundation (DNRF grant No. 141 to the Center for Functional Genomics and Tissue Plasticity [ATLAS]) and grants from the Novo Nordisk Foundation through a grant to the Danish Diabetes Academy. The authors thank Tenna Pavia Mortensen for expert technical assistance and colleagues from the Functional Genomics & Metabolism Research Unit for fruitful discussions. Methodology, E.L.V.H. E.G. and L.L.; investigation, E.L.V.H. E.G. A.K.S. L.L. and R.N.; validation, E.L.V.H. E.G. L.L.; writing ? original draft, E.L.V.H. and S.M.; writing ? review & editing, S.M. J.G.S.M. E.L.V.H. E.G. and A.K.S.; visualization, E.L.V.H. and E.G.; supervision, S.M. and J.G.S.M.; project administration, S.M. and J.G.S.M.; funding acquisition, S.M. and E.G. The authors declare no competing interests.

Publisher Copyright:
© 2021 The Author(s)

Dokumenter og links

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

Open Access VersionForlagets udgivne version, 2,61 MBLicens: CC BY-NC-ND

1 Ph.d.-afhandling

Adipose tissue plasticity in obesity
Bidragets oversatte titel: Plasticitet af fedtvæv i fedmeHauwaert, E. L. V., 10. feb. 2022, Syddansk Universitet. Det Naturvidenskabelige Fakultet. 99 s.
Publikation: Afhandling › Ph.d.-afhandling

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Citationsformater

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AU - Larsen, Lena

AU - Nielsen, Ronni

AU - Madsen, Jesper Grud Skat

AU - Mandrup, Susanne

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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.

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KW - Gene Expression

KW - Genomics

KW - Metabolism

KW - RNA-seq

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

Abstract

Bibliografisk note

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Adipose tissue plasticity in obesity

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