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

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...

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Detalles Bibliográficos
Autores principales: Van Hauwaert, Elvira Laila, Gammelmark, Ellen, Sárvári, Anitta Kinga, Larsen, Lena, Nielsen, Ronni, Madsen, Jesper Grud Skat, Mandrup, Susanne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8220393/
https://www.ncbi.nlm.nih.gov/pubmed/34189477
http://dx.doi.org/10.1016/j.xpro.2021.100612
Descripción
Sumario: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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