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Ex vivo proliferation of mesothelial cells directly obtained from peritoneal effluent: its relationship with
1Hospital Universitario La Paz, Madrid, Spain.
Abstract:
The peritoneal membrane requires anatomico-functional integrity to guarantee long-term stability for peritoneal dialysis (PD). Since mesothelial cells (MC) are active cells and the first part of the membrane to contact the dialysate, they are important in maintaining this stability. Mesothelial cells released daily into peritoneal effluent are able to grow in culture. This growth capacity may be related to some of the anatomicofunctional characteristics of each peritoneum. Our aim was to culture mesothelial cells taken from peritoneal effluents drained by 32 PD-stable patients, and relate this growth capacity to individual peritoneal data. Cells were taken from a residual fluid after sedimentation, washed twice with phosphate-buffered saline (PBS), and seeded into 25-cm2 tissue-culture flasks. These flasks were incubated in a humidified 5%-CO2 atmosphere. After MC confluence, cells were detached by trypsinization, passaged into 24-well plates, and finally counted. Cells were identified by morphology and immuno-histochemical characteristics. Cells from 28 out of 32 patients showed an appropriate growth in culture. Mesothelial cell confluence was reached in a mean of 18.2 +/- 8 days. After 7 days of seeding in plate wells, the cell growth showed a significant and progressive increase until day 16. Mesothelial cell growth rate was inversely related to PD duration. Neither peritonitis incidence nor other demographic characteristic were related to MC growth. Creatinine and urea mass transfer coefficients (MTC), but not ultrafiltration (UF) capacity, were significantly related to MC growth rate. In conclusion, the growth in culture of MC taken directly from PD bags is certainly possible. This growth is influenced by some of the intrinsic peritoneal characteristics derived from the peritoneal dialysis process. This tool could be useful in evaluating individual peritoneal conditions and, probably, as a method for peritoneal viability follow-up, although further research is required.
Insights
Culturing mesothelial cells (MC) from peritoneal dialysis (PD) effluent is feasible. MC growth capacity in vitro correlates with peritoneal membrane function, offering potential for assessing patient dialysis status.
Area of Science:
- Nephrology
- Cell Biology
- Biomaterials Science
Background:
- Peritoneal dialysis (PD) relies on the peritoneal membrane's integrity for long-term success.
- Mesothelial cells (MC) are crucial for maintaining peritoneal membrane stability during PD.
- MC shed into peritoneal effluent retain growth potential in culture.
Purpose of the Study:
- To culture mesothelial cells (MC) from peritoneal dialysis (PD) effluent of stable patients.
- To investigate the relationship between MC growth capacity and individual peritoneal functional data.
Main Methods:
- MC were isolated from residual fluid in PD effluent bags after sedimentation.
- Cells were cultured, passaged, and growth rates were assessed.
- Cell morphology and immuno-histochemical markers confirmed MC identity.
Main Results:
- Successful culture and growth of MC were achieved in 28 out of 32 patients.
- MC confluence was reached in a mean of 18.2 days, with significant growth observed up to day 16.
- MC growth rate showed an inverse relationship with PD duration and correlated significantly with creatinine and urea mass transfer coefficients (MTC).
Conclusions:
- Culturing mesothelial cells (MC) from PD effluent is a viable method.
- MC growth in vitro reflects intrinsic peritoneal characteristics influenced by PD.
- This technique may serve as a tool for evaluating individual peritoneal conditions and monitoring viability.