Author/Editor		Burja, Blaž; Paul, Dominique; Tastanova, Aizhan; Edalat, Sam G.; Sodin-Šemrl, Snežna; Lakota, Katja; Tomšič, Matija; Rotar, Žiga; Frank Bertoncelj, Mojca
Title		An optimized tissue dissociation protocol for single-cell rna sequencing analysis of fresh and cultured human skin biopsies
Type		članek
Vol. and No.		, št. Vol.
Publication year		2022
Volume		str. str.
ISSN		2296-634X - Frontiers in cell and developmental biology (Online)
Language		eng
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 ex vivo 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 ex vivo skin explant experimentations.
Keywords		skin tissue skin biopsy scRNAseq kožno tkivo biopsija kože scRNAseq