Isolation of nuclei from frozen human subcutaneous adipose tissue for full-length single-nuclei transcriptional profiling

Automated single-cell dispensing is incompatible with white adipose tissue (WAT) due to lipid-laden adipocytes. Single-nuclei RNA-Seq permits transcriptional profiling of all cells from WAT. Human WAT faces unique technical challenges in isolating nuclei compared to rodent tissue due to greater extr...

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Detalles Bibliográficos
Autores principales: Whytock, Katie L., Divoux, Adeline, Sun, Yifei, Hopf, Meghan, Yeo, Reichelle X., Pino, Maria F., Yu, GongXin, Smith, Steven R., Walsh, Martin J., Sparks, Lauren M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9876942/
https://www.ncbi.nlm.nih.gov/pubmed/36853719
http://dx.doi.org/10.1016/j.xpro.2023.102054
Descripción
Sumario:Automated single-cell dispensing is incompatible with white adipose tissue (WAT) due to lipid-laden adipocytes. Single-nuclei RNA-Seq permits transcriptional profiling of all cells from WAT. Human WAT faces unique technical challenges in isolating nuclei compared to rodent tissue due to greater extra-cellular matrix content and larger lipid droplets. In this protocol, we detail how to isolate nuclei from frozen subcutaneous human WAT for single-nuclei RNA-Seq. For complete information on the generation and use of this protocol, please refer to Whytock et al. (2022).(1)

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