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

Katie L. Whytock, Adeline Divoux, Yifei Sun, Meghan Hopf, Reichelle X. Yeo, Maria F. Pino, Gong Xin Yu, Steven R. Smith, Martin J. Walsh, Lauren M. Sparks

Research output: Contribution to journalArticlepeer-review

Abstract

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

Original languageEnglish
Article number102054
JournalSTAR Protocols
Volume4
Issue number1
DOIs
StatePublished - 17 Mar 2023

Keywords

  • Cell isolation
  • Metabolism
  • RNAseq
  • Single Cell

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