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

Abstract

Summary: 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
• Single Cell
• Genomics
• RNA-seq
• Metabolism
• Gene Expression