Single-cell RNA sequencing is a promising and robust technique that has been widely applied in the field of dermatology to study the transcriptional profile of each cell at very high resolution. Here we tested the compatibility of the novel instrument-free single-cell RNA-sequencing kit, recently developed by Scipio bioscience, on human skin cells. Human skin punch biopsy was first mechanically treated for a separation into dermis and epidermis fractions, then each fraction was separately processed for cell dissociation. Subsequently, single-cell RNA sample preparation was achieved using the proprietary reversible hydrogel technology that provides results comparable to current microfluidic instrument-based techniques. In the application reported here, chemically linked barcode bead—cell tandems were dispersed in the hydrogel in the liquid state. Upon gelation the tandems were immobilized, but small lysis agents that diffuse freely were subsequently added. Following cell lysis mRNA molecules from individual cells were captured on the adjacent barcodes. Successive steps were reverse transcription, PCR amplification, cDNA sequencing, and bioinformatic analysis. We profiled the transcriptome of 3609 single cells from healthy skin. We identified 15 cell subtypes in the epidermis and 15 cell subtypes in the dermis. SingleR annotation based on reference data revealed all the major cell types commonly described in the literature as well as rare cell populations such as T-cells, dendritic cells, macrophages and Langerhans cells. Furthermore, the gene expression signatures were assessed and were comparable to the data from literature. We demonstrate single-cell RNA-sequencing using Scipio bioscience’s kit as an effective strategy to dissect the heterogeneity and the complexity of human skin.

ESDR279 - Instrument-free single-cell resolution of transcriptome in human skin

The scientific meeting of the European Society for Dermatological Research (ESDR) is held yearly, usually in September. With an international attendance of more than 1000 skin scientists from around the globe, the four-day event includes short oral presentations from peer-reviewed abstracts, keynote lectures from international experts, sponsored symposia, satellite programs, educational events as well as an important focus on posters, networking and building collaborations amongst scientists. For more information about this year’s meeting visit our website.