GSM8425828: GMC01, scRNAseq; Homo sapiens; RNA-Seq
After collection, fresh fascial samples were stored at 4 °C in the GEXSCOPE Tissue Preservation Solution (Cat# 1200050001, Singleron Biotechnologies, Nanjing, China) and processed within 24 h. The samples underwent three washes with Hanks Balanced Salt Solution and were then minced into small pieces measuring 1–2 mm3. Subsequently, they were digested in GEXSCOPE Tissue Dissociation Mix Buffer Solution (Cat# 1200000044, Singleron Biotechnologies, Nanjing, China) inside a thermostatic oscillator at 37 °C and 180 rpm for 15 min. Tissue lysates were filtered through a 40 µm sterile strainer (Cat# 352340, Corning, New York, USA), and any remaining red blood cells were eliminated by incubating in Red Blood Cell Lysis Buffer (Cat# 00‑4333‑57, ThermoFisher, Waltham, USA) at 4 °C for 10 min. The viability of finial single‑cell suspensions was determined using Trypan Blue (Cat# T8154, Sigma, St. Louis, USA) and acridine orange/propidium iodide staining (Cat# CS2‑0106, Nexcelom, Lawrence, USA). ScRNA‑seq libraries were constructed according to the manufacturer's protocol (GEXSCOPE Single Cell RNA seq Kit, Singleron Biotechnologies, Nanjing, China). The resulting scRNA‑seq libraries were sequenced on the Illumina NovaSeq 6000 platform with 150 bp paired‑end reads, aiming for a sequence depth of approximately 30,000 reads per cell.