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.
Bibliografisk noteFunding Information:
The work was supported by grants from the Danish National Research Foundation (DNRF grant No. 141 to the Center for Functional Genomics and Tissue Plasticity [ATLAS]) and grants from the Novo Nordisk Foundation through a grant to the Danish Diabetes Academy. The authors thank Tenna Pavia Mortensen for expert technical assistance and colleagues from the Functional Genomics & Metabolism Research Unit for fruitful discussions. Methodology, E.L.V.H. E.G. and L.L.; investigation, E.L.V.H. E.G. A.K.S. L.L. and R.N.; validation, E.L.V.H. E.G. L.L.; writing ? original draft, E.L.V.H. and S.M.; writing ? review & editing, S.M. J.G.S.M. E.L.V.H. E.G. and A.K.S.; visualization, E.L.V.H. and E.G.; supervision, S.M. and J.G.S.M.; project administration, S.M. and J.G.S.M.; funding acquisition, S.M. and E.G. The authors declare no competing interests.
© 2021 The Author(s)