scRNA-seq performed in the knee joint of the DMM-induced OA mouse model versus sham-operated mice

Osteoarthritis (OA) is typically characterized by progressive cartilage breakdown, subchondral bone remodeling, osteophyte formation and low-grade joint inflammation. Accumulative evidence demonstrates that cartilage fibrosis and the resulting degradation of extracellular matrix are implicated in the late-stage pathology of OA. However, the mechanisms underlying this degenerative joint disease remain largely undefined.We isolated articular cartilage from control-sham, control-DMM and cKOAcan(chondrocyte conditional ZEB1 knockout mice)-DMM mice and performed scRNA-seq analysis. Overall design: The cartilage from control-sham, control-DMM and cKOAcan-DMM mice was collected and stored in the sCelLiveTM Tissue Preservation Solution (Singleron) on ice within 30 mins. After being washed with Hank's Balanced Salt Solution, the samples were cut into 1-2mm pieces and then digested with 3 mL sCelLiveTM Tissue Dissociation Solution (Singleron) by Singleron PythoN™ Tissue Dissociation System at 37°C for 15 min. The cell suspension was filtered through a 40-micron sterile strainer to remove red blood cells and stained with Trypan Blue to evaluate the cell viability. The mRNA from single cells was reversely transcribed into cDNA and the cDNA was amplified by PCR, then fragmented and ligated with sequencing adapters. The GEXSCOPE® Single Cell RNA Library Kit (Singleron) was used to construct the scRNA-seq libraries. CeleScope v1.5.2 (Singleron Biotechnologies) was used to generate gene expression profiles with default parameters and the raw data was used for further analysis.