Single cell RNA sequencing analysis of the center, edge and non-wounded regions of healing skin on day 11 post-wounding in wild type mice

We applied single-cell RNA sequencing to investigate specific type of cell populations and their gene expressions on the day of wound closure (post-wounding day 11-12) at the center and edge of the re-epithelialized mouse wound site (1 cm² of whole skin was removed, with samples pooled from 10 mice). Skin samples from non-wounded area were collected as control. Overall design: Viable cells from the single-cell suspension were selected by sorting, and a single-cell library was prepared using 10x Genomics and sequenced with the Illumina NovaSeq 6000. Downstream bioinformatic analysis of single-cell RNA sequencing data was performed using the Seurat package in R. Cell clusters were annotated according to their identity using specific marker genes.

我们采用单细胞RNA测序（single-cell RNA sequencing），在伤口闭合当日（伤后第11-12天）对再上皮化小鼠伤口创面的中心与边缘区域的特定细胞群及其基因表达特征展开探究。本次实验移除了1平方厘米的全层皮肤，样本由10只小鼠的组织混合制备；同时收集未受伤区域的皮肤样本作为对照。实验整体设计：通过分选获取单细胞悬液中的活细胞，利用10x Genomics平台构建单细胞文库，并使用Illumina NovaSeq 6000完成测序。单细胞RNA测序数据的下游生物信息学分析依托R语言中的Seurat软件包完成；细胞簇通过特异性标记基因的表达特征完成身份注释。