Single cell RNA sequencing reveals dynamic changes in the peritoneal microenvironment of patients undergoing dialysis for 3 years and provides new insights for peritoneal fibrosis

Peritoneal dialysis related fibrosis (PD-related fibrosis) is one of the most common complications of PD and one of the main reasons for patients to stop PD, occurring almost exclusively in all long-term PD patients. The mechanism of PD-related fibrosis is currently not fully understood. Previous studies have shown that it is related to frequent peritonitis and the epithelial-mesenchymal transition (more accurately, mesothelial-mesenchymal transition) of peritoneal mesothelial cells caused by high glucose and high osmotic non physiologically compatible PD fluid. Due to the lack of effective prevention and treatment methods for PD-related fibrosis, the study of its mechanism is of great clinical significance. Collecting dialysis fluid from patients after 3 years of PDfor single-cell sequencing can help observe changes in the peritoneal environment and structural composition during dialysis, providing a new perspective for the prevention and treatment of PD-related fibrosis. Overall design: We collected 2 liters of overnight PD fluid from a patient who underwent continuous ambulatory peritoneal dialysis (CAPD) for 3 years. Dialysis fluid is obtained within 1 hour after the patient's dialysis fluid flows out. Obtain exfoliated cells from dialysate by centrifugation at 3000rpm for 15 minutes, wash the cells twice with phosphate buffered saline (PBS) buffer, and finally resuspend the cells in culture medium containing 10% bovine serum albumin (BSA).