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

STAR Protoc. 2021 Jun 17;2(3):100612. doi: 10.1016/j.xpro.2021.100612. eCollection 2021 Sep 17.

Authors

Elvira Laila Van Hauwaert¹, Ellen Gammelmark¹, Anitta Kinga Sárvári¹, Lena Larsen¹, Ronni Nielsen¹, Jesper Grud Skat Madsen¹, Susanne Mandrup¹

Affiliation

¹ Center for Functional Genomics and Tissue Plasticity, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M 5230, Denmark.

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.

Keywords: Cell isolation; Gene Expression; Genomics; Metabolism; RNA-seq; Single Cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adipose Tissue, White / metabolism*
Animals
Cell Nucleus / metabolism*
Genomics*
Mice
Mice, Inbred C57BL
Single-Cell Analysis / methods