AbstractThe skin harbours transcriptionally and functionally heterogeneous mesenchymal cells that participate in various physiological activities by secreting biochemical cues. In this study, we aimed to identify a new subpopulation of dermal mesenchymal cells that enhance hair follicle regeneration through a paracrine mechanism. Integrated single‐cell RNA sequencing (scRNA‐seq) data analysis revealed epidermal growth factor receptor (EGFR) as a marker of distinct fibroblast subpopulation in the neonatal murine dermis. Immunofluorescence staining and fluorescence‐activated cell sorting (FACS) were used to validate the existence of the cell population inKrt14‐rtTA‐H2BGFPmouse. The difference of gene expression between separated cell subpopulation was examined by real‐time PCR. Potential effect of the designated factor on hair follicle regeneration was observed after the application on excisional wounds inKrt14‐rtTA‐H2BGFPmouse. Immunofluorescence staining demonstrated the existence of dermal EGFR+cells in neonatal and adult mouse dermis. The EGFR+mesenchymal population, sorted by FACS, displayed a higher expression level ofIgf1(insulin‐like growth factor 1). Co‐localisation of IGF1 with EGFR in the mouse dermis and upregulated numbers of hair follicles in healed wounds following the application of exogenous IGF1 illustrated the contribution of EGFR+cells in promoting wound‐induced hair follicle neogenesis. Our results indicate that EGFR identifies a subpopulation of dermal fibroblasts that contribute to IGF1 promotion of hair follicle neogenesis. It broadens the understanding of heterogeneity and the mesenchymal cell function in skin and may facilitate the potential translational application of these cells.