GSM9237580: 4M; Homo sapiens; RNA-Seq
The isolated TMJC tissue was cut into 2–3 mm sections and rinsed with Hanks' Balanced Salt Solution (HBSS). Enzymatic digestion was performed for 15 minutes using a tissue dissociation solution (Singleron Biotechnologies, Nanjing, China). The resulting cell suspension was filtered through a sterile 40-μm filter (Corning, NY, USA) and centrifuged at 150 g for 5 minutes. Erythrocytes were removed with a lysis reagent (Singleron Biotechnologies, Nanjing, China) for 10 minutes. The cells were resuspended in phosphate-buffered saline (PBS) and stained with trypan blue (T6146, Sigma, Burlington, VT, USA). Cell viability was assessed using a TC20 automated cell counter (Bio-Rad, Hercules, CA, USA). A cell suspension at a concentration of 1×10^5 cells/mL was prepared and loaded into a microfluidic plate. The scRNA-seq libraries were constructed using the Single-Cell RNA Library Kit (Singleron Biotechnologies, Nanjing, China). The prepared libraries were pooled and sequenced on an Illumina NovaSeq 6000 platform. Raw sequencing reads were processed with the CeleScope pipeline (https://github.com/singleron-RD/CeleScope).