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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 Sárvári et al., 2021.

SUBMITTER: Van Hauwaert EL

PROVIDER: S-EPMC8220393 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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

Van Hauwaert Elvira Laila EL Gammelmark Ellen E Sárvári Anitta Kinga AK Larsen Lena L Nielsen Ronni R Madsen Jesper Grud Skat JGS Mandrup Susanne S

STAR protocols 20210617 3

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 ...[more]

PMID: 34189477

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