RNA Sequencing of Porcine Osteoarthritic Chondrocytes to Investigate Cartilage Pathophysiology
Porcine chondrocytes were isolated from full-depth cartilage obtained from porcine metacarpophalangeal joints through sequential enzymatic digestion using pronase and collagenase. The isolated cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 15% fetal bovine serum (FBS), 1% penicillin-streptomycin, 10 mM HEPES, 4 mM L-glutamine, and 0.15 mg/mL L-ascorbic acid. After 24 hours of culture without passage, total RNA was extracted using Trizol Reagent. RNA sequencing (RNA-seq) was performed by Gene Denovo Biotechnology Co. (Guangzhou, China) on the Illumina HiSeq2500 platform.