Single-cell analysis reveals the progress in skin repair after radiation in rats.

This study aims to compare the cell heterogeneity and mRNA expression between irradiated and non-irradiated rat skin tissues at the single cell level. In this study, the skin was isolated from rats with radiation or non-irradiated rats. Male Sprague Dawley (SD) rats (6 weeks of age) were used. For irradiation, the hair on the gluteal region of the rats was shaved using a razor. The animals were immobilized using adhesive tape on a plastic plate to minimize motion during radiation exposure. A 3-cm-thick piece of lead was used to shield the animals and localize the radiation field (3-4 cm). Irradiation was performed on the treatment area at a dose rate of 1000 cGy/min using a 6-MeV electron beam accelerator. Irradiated skin was taken on day 7, 14, 28 and 60 for RNA-seq, respectively. The control group of skin were collected from four rats without radiation and mixed for RNA-seq. Overall design: Single cell RNA-seq analysis of skin tissue from irradiated and non-irradiated parts of SD rats at different time periods. ScRNA sequencing libraries were further generated by the 10x Chromium Single Cell 3' Library & Single Cell 3' v3 Gel Beads (10x Genomics). Briefly, gel beads with cell barcodes and skin cell suspension were wrapped in liquid droplets by a microfluidic platform (10x Genomics). A droplet containing single cells was collected and then lysed to generate Gel Bead-In Emulsions (GEMs).