21.09.2021
Van Hauwaert et al. present an optimized protocol for isolating nuclei from mouse adipose tissue to enable single-nucleus RNA sequencing of lipid-filled adipocytes. They demonstrate that this approach overcomes limitations of droplet-based single-cell methods and allows comprehensive transcriptomic profiling of whole adipose tissue.
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.
https://doi.org/10.1016/j.xpro.2021.100612
