Processing human skin samples for single-cell assays

ABSTRACT: Summary Characterizing resident immune cells in human skin using single-cell assays provides insight into their role in infections, inflammation, and cancer. We describe an optimized protocol to rapidly isolate viable cells from 6-mm skin punch-biopsies. We provide an example in which we coupled single-cell RNA sequencing (scRNA-seq) with single-cell T-cell receptor sequencing (scTCR-seq) of skin and blood cells to study transcriptional profiles and clonotypes of skin resident and peripheral circulating, memory, and effector T cells. This is an improved protocol based on Saluzzo et al. (2021). For complete details on the use and execution of this protocol, please refer to Saluzzo et al. (2021). Graphical abstract Highlights • Preparation of a high-quality single-cell suspension from human skin punch biopsies• Suitable for single-cell RNA and TCR sequencing to study skin and blood T cells• Cost minimization by hash-tagging and pooling blood and skin cells of one individual• Overview of the downstream bioinformatic analysis for the T cell characterization Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Characterizing resident immune cells in human skin using single-cell assays provides insight into their role in infections, inflammation, and cancer. We describe an optimized protocol to rapidly isolate viable cells from 6-mm skin punch-biopsies. We provide an example in which we coupled single-cell RNA sequencing (scRNA-Seq) with single-cell T-cell receptor sequencing (scTCR-Seq) of skin and blood cells to study transcriptional profiles and clonotypes of skin resident and peripheral circulating, memory, and effector T cells. This is an improved protocol based on Saluzzo et al. (2021).

SUBMITTER: Saluzzo S 

PROVIDER: S-EPMC9234085 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature