Paracrine effects of transplanted mesothelial cells isolated from temperature-sensitive SV40 large T-antigen gene transgenic rats during peritoneal repair

Abstract

BackgroundThe prevention and restoration of peritoneal damage is a critical mission in peritoneal dialysis (PD). Transplantation of mesothelial cells has been suggested to suppress peritoneal injury during PD. Few studies have examined the efficacy and safety of cell transplantation. We evaluated the paracrine effects of mesothelial transplantation during peritoneal repair using immortalized temperature-sensitive mesothelial cells (TSMCs) in chlorhexidine gluconate (CG)-induced peritoneal fibrosis rats.MethodsContinuous-infusion pumps containing 8% CG were placed into the abdominal cavity for 21 days. After the removal of the pumps, the TSMCs were injected into the peritoneal cavity at Day 22 (Tx-1 group) or 29 (Tx-2 group). Morphological findings and mRNA expressions of regeneration-related factors were examined at Days 22, 29 and 35.ResultsPeritoneal thickness was aggravated in the Tx-1 group. Levels of transforming growth factor (TGF)-β, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 mRNA in the Tx-1 group at Day 35 were comparable with those at Day 22. The levels of Snail, B-Raf and ERK-1, markers of epithelial to mesenchymal transition and of the RAS/MAPK pathway in the Tx-1 group, were significantly higher than those in the Tx-2 group. TGF-β and VEGF were produced from the transplanted mesothelial cells and the surrounding cells in the Tx-1 group.ConclusionIt appears that the paracrine effect of transplanted mesothelial cells during peritoneal repair is associated with its surrounding condition. It is important to determine the most appropriate time for developing peritoneal repair through mesothelial transplantation.

Adipose-derived mesenchymal stem cells transplantation facilitate experimental peritoneal fibrosis repair by suppressing epithelial–mesenchymal transition

“…Animal models of peritoneal fibrosis Rats were continuously infused with 8 % CG using infusion pumps as previously described [21,22]. In brief, infusion pumps were placed in the abdominal cavity for 21 days and the anterior abdominal peritoneum was harvested on days 0, 22, 29 and 35 (CG group).…”

“…Immunohistochemical analysis was performed using formalin fixed tissue sections after microwave antigen retrieval as previously described [21]. The mouse anti-rat alphasmooth muscle actin (a-SMA) antibody (Abcam, Cambridge, UK) was used as primary antibody.…”

“…Immunofluorescence was performed using formalin fixed tissue sections after microwave antigen retrieval as previously described [21]. Sections were incubated overnight with the following antibodies: fluorescein isothiocyanate (FITC)-conjugated goat anti-GFP antibody (Abcam, Tokyo, Japan), mouse anti-VEGF antibody (Abcam), mouse anti-CD31 antibody (Abcam), rabbit anti-plateletderived growth factor-BB (PDGF-BB) antibody (Abcam), rabbit anti-PDGF-Rb antibody (Abcam), and rabbit anti-a-SMA antibody (Abcam).…”

“…RNA extraction and real-time PCR analysis were performed as previously described [21]. For quantification, samples were normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA.…”

Peritonitis-induced peritoneal injury models for research in peritoneal dialysis review of infectious and non-infectious models

“…Peritoneal resting for 3 weeks after 3 weeks of CG exposure ameliorated some functional parameters in the peritoneum; however, elevated peritoneal thickness and fibrosis continued during the resting period [88][89][90][91]. Placing an infusion pump in the rat abdominal cavity was reported as an alternative administration route for CG [92][93][94].…”