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

Research output: Contribution to journalJournal articleResearchpeer-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 languageEnglish
Article number100612
JournalSTAR Protocols
Volume2
Issue number3
Number of pages12
DOIs
Publication statusPublished - 17. Sep 2021

Bibliographical note

Publisher Copyright:
© 2021 The Author(s)

Keywords

  • Cell isolation
  • Gene Expression
  • Genomics
  • Metabolism
  • RNA-seq
  • Single Cell

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