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; GongXin Yu; Steven R Smith; Martin J Walsh; Lauren M Sparks

doi:10.1016/j.xpro.2023.102054

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

STAR Protoc. 2023 Mar 17;4(1):102054. doi: 10.1016/j.xpro.2023.102054. Epub 2023 Jan 20.

Authors

Affiliations

¹ Translational Research Institute, AdventHealth, Orlando, FL 32804, USA. Electronic address: katie.whytock@adventhealth.com.
² Translational Research Institute, AdventHealth, Orlando, FL 32804, USA.
³ Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
⁴ Translational Research Institute, AdventHealth, Orlando, FL 32804, USA. Electronic address: lauren.sparks@adventhealth.com.

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.

Trial registration: ClinicalTrials.gov NCT04034706.

Keywords: Cell isolation; Metabolism; RNAseq; Single Cell.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Adipocytes
Adipose Tissue, White*
Cell Nucleus / genetics
Humans
RNA-Seq
Subcutaneous Fat*

Associated data

ClinicalTrials.gov/NCT04034706

Abstract

Publication types

MeSH terms

Associated data

Grants and funding