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

从小鼠白色脂肪组织中分离细胞核用于单核基因组学研究

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

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.

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