title: Distinct diversity of skin cell populations of rhinophyma and hypertrophic scar illustrated by scRNA-seq
design: Distinct diversity of skin cell populations of rhinophyma and hypertrophic scar illustrated by scRNA-seq
library_construction_protocol: This study was approved by the Ethics Committee of Shuguang Hospital (No.2024-1488-071-01 and No.2024-1598-181-01). Written informed consent was obtained from all participants. Hypertrophic scar tissue was collected during elective plastic surgery. Rhinophyma specimens were harvested via five-blade scarification surgery. Normal skin was acquired from donors undergoing routine circumcision. Each sample measured approximately 0.5 cm³ (normal skin: 0.25 cm³, two samples per donor). Patients had received no prior topical/systemic treatments affecting the sampled areas for ≥4 weeks before surgery. All procedures complied with the Declaration of Helsinki. Tissues were minced and enzymatically digested with 0.75mg/mL collagenase I, 2mg/mL collagenase IV, 0.2mg/ml hyaluronidase and 75 U/mL DNase I for 45 min at 37°C. Cells were filtered through 70µm and 35µm strainers, centrifuged, and resuspended. Cell viability was assessed using AO/PI staining and Countstar Fluorescence Cell Analyzer. Single-cell RNA-seq libraries were generated using the 10X Genomics Chromium Controller Instrument and Chromium Single Cell 3' V3.1 Reagent Kits. Cells were loaded to generate Gel Bead-In-Emulsions (GEMs). After reverse transcription, barcoded cDNA was amplified, fragmented, and ligated with adaptors. Library quality and quantity were assessed using the Qubit High Sensitivity DNA assay and Bioanalyzer 2200.