GSM9814425: Patient 1 Hypercellular zone; Homo sapiens; RNA-Seq
issue dissociation and single-cell suspension preparation: Keloid samples were preserved in GEXSCOPE tissue preservation solution and transferred to the laboratory under cold conditions. Samples were washed three times with Hanks Balanced Salt Solution (HBSS), cut into 1-2 mm fragments, and digested in 2 ml GEXSCOPE tissue dissociation solution (Singleron Bio, Nanjing, China). After digestion, the mixture was filtered, centrifuged, and the pellet resuspended in PBS. Cell viability and concentration were assessed using trypan blue staining and Countstar Rigel fluorescence cell counter. Single-cell RNA library construction and sequencing: Single-cell suspensions (2×10^5 cells/mL) were loaded onto microwell chips using the Singleron Matrix Single Cell Processing System. scRNA-seq libraries were constructed with GEXSCOPE Single Cell RNA Library Kits (Singleron) following the manufacturer's protocol. Libraries were sequenced on Illumina NovaSeq 6000 with 150 bp paired-end reads. Single-cell suspensions (2×10⁵ cells/mL in PBS) were loaded onto microwell chips using the Singleron Matrix® Single Cell Processing System. scRNA-seq libraries were constructed according to the manufacturer's protocol using the GEXSCOPE® Single Cell RNA Library Kits (Singleron Bio, Nanjing, China). Individual libraries were quantified, diluted to 4 nM, pooled, and sequenced on an Illumina NovaSeq 6000 platform with 150 bp paired-end reads.