title: GSM8073280: IVD, scRNA seq; Mus musculus; RNA-Seq
library_construction_protocol: NP tissue was meticulously extracted from the lumbar and tail IVDs under a microscope, digested with 0.1% pronase (Sigma-Aldrich, Burlington, MA, USA) at 37°C for 30 minutes, and further digested with 0.2% collagenase type 2 (Sigma-Aldrich, Burlington, MA, USA) at 37°C for 2 hours.Then CD45+ immune cells were removed by FACS wtih APC/Cyanine7 anti-mouse CD45 Antibody(Biolegend, USA). Approximately 16,000 cells were loaded into microfluidic chips to generate single-cell gel beads in emulsion by using a commercial kit (Chromium Next GEM Single Cell 5ʹ GEM Kit v2, 10X Genomic, Pleasanton, USA,). Subsequent cDNA amplification, quality control and library construction were carried out in accordance with standard guidelines.