Isolation of Human Cutaneous Immune Cells for Single-Cell RNA Sequencing

Authors

Ashley A Hailer
David Wu
Abdullah El Kurdi
Michelle Yuan
Raymond J Cho
Jeffrey B Cheng

Language

English

Publication Date

4-28-2023

Journal

STAR Protocols

DOI

10.1016/j.xpro.2023.102239

PMID

37120815

PMCID

PMC10173011

PubMedCentral® Posted Date

4-28-2023

PubMedCentral® Full Text Version

Post-print

Abstract

Single-cell RNA sequencing (scRNA-seq) allows for high-resolution analysis of transcriptionally dysregulated cell subpopulations in inflammatory diseases. However, it can be challenging to properly isolate viable immune cells from human skin for scRNA-seq due to its barrier properties. Here, we present a protocol to isolate high-viability human cutaneous immune cells. We describe steps for obtaining and enzymatically dissociating a skin biopsy specimen and isolating immune cells using flow cytometry. We then provide an overview of downstream computational techniques to analyze sequencing data. For complete details on the use and execution of this protocol, please refer to Cook et al. (2022)

Keywords

Cell Isolation, Flow Cytometry/Mass Cytometry, Immunology, Molecular Biology, RNAseq, Single Cell

Published Open-Access

yes

Recommended Citation

Hailer, Ashley A; Wu, David; El Kurdi, Abdullah; et al., "Isolation of Human Cutaneous Immune Cells for Single-Cell RNA Sequencing" (2023). Staff and Researcher Publications. 23.
https://digitalcommons.library.tmc.edu/clinic_pub/23