An Optimized Tissue Dissociation Protocol for Single-Cell RNA Sequencing Analysis of Fresh and Cultured Human Skin Biopsies

Overview

Journal Front Cell Dev Biol

Specialty Cell Biology

Date 2022 May 16

PMID 35573685

Authors

Blaz Burja

Dominique Paul

Aizhan Tastanova

Sam G Edalat

Reto Gerber

Miranda Houtman

Muriel Elhai

Kristina Burki

Ramon Staeger

Gaetana Restivo

Ramon Lang

Snezna Sodin-Semrl

Katja Lakota

Matija Tomsic

Mitchell P Levesque

Oliver Distler

Ziga Rotar

Mark D Robinson

Mojca Frank-Bertoncelj

Affiliations

Soon will be listed here.

Abstract

We present an optimized dissociation protocol for preparing high-quality skin cell suspensions for in-depth single-cell RNA-sequencing (scRNA-seq) analysis of fresh and cultured human skin. Our protocol enabled the isolation of a consistently high number of highly viable skin cells from small freshly dissociated punch skin biopsies, which we use for scRNA-seq studies. We recapitulated not only the main cell populations of existing single-cell skin atlases, but also identified rare cell populations, such as mast cells. Furthermore, we effectively isolated highly viable single cells from cultured skin biopsy fragments and generated a global single-cell map of the explanted human skin. The quality metrics of the generated scRNA-seq datasets were comparable between freshly dissociated and cultured skin. Overall, by enabling efficient cell isolation and comprehensive cell mapping, our skin dissociation-scRNA-seq workflow can greatly facilitate scRNA-seq discoveries across diverse human skin pathologies and skin explant experimentations.

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