title: Ex vivo proliferation of mesothelial cells directly obtained from peritoneal effluent: its relationship with peritoneal antecedents and functional parameters.
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.
authors.affiliation: Hospital Universitario La Paz, Madrid, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Díaz
authors.forename: R
authors.initials: R
authors.lastname: Selgas
authors.forename: M A
authors.initials: MA
authors.lastname: Castro
authors.forename: M A
authors.initials: MA
authors.lastname: Bajo
authors.forename: M
authors.initials: M
authors.lastname: Fernández de Castro
authors.forename: S
authors.initials: S
authors.lastname: Molina
authors.forename: C
authors.initials: C
authors.lastname: Jiménez
authors.forename: A
authors.initials: A
authors.lastname: Ortiz
authors.forename: F
authors.initials: F
authors.lastname: Vara
mesh_terms: D001202:Ascitic Fluid / Q000166:cytology*; D002455:Cell Division; D002478:Cells, Cultured; D003404:Creatinine / Q000378:metabolism; D015314:Dialysis Solutions; D004847:Epithelial Cells / Q000166:cytology*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D010530:Peritoneal Dialysis; D010537:Peritoneum / Q000166:cytology / Q000378:metabolism; D014508:Urea / Q000378:metabolism
chemical_list: D015314:Dialysis Solutions; D014508:Urea; D003404:Creatinine