In vitro of the effect of osmotic solutes on the interactions between cells from the peritoneum and peritoneal cavity

Objective: To study how the presence of osmotic solutes in medium affects growth of the peritoneal mesothelial cells and fibroblasts and how osmotic solutes influence the production of factors regulating growth of these cells. Design: The proliferation of mesothelial cells and fibroblasts was evaluated by measuring the incorporation of 3H-thymidine into the cells. Cells were exposed to osmotic solutes; the concentration of the latter in the medium was continuously lowered over the time of the experiment to simulate changes of their concentration in the dialysate. The synthesis of factors influencing the proliferation of the mesothelial cells or fibroblasts, by mesothelial cells or fibroblasts themselves, or by peritoneal leukocytes, was tested by the characteristics of the 'conditioned' medium. The conditional medium was produced by exposing standard medium to mesothelial or fibroblasts monolayer or to peritoneal leukocytes over 24 hours; following it was applied to growing test cells for the study of growth factors. Results: The effect of osmotic solutes on the growth of mesothelial cells is less inhibitory when their concentration is gradually lowered over the time of the study, compared to previous findings with a constant concentration. Peritoneal leukocytes produce growth factors for mesothelial cells and fibroblasts. Glucose and amino acids inhibit production of peritoneal leukocyte-derived growth factors for mesothelial cells, while glycerol increases synthesis of such growth factors for fibroblasts. Mesothelial cells produce factors stimulating the proliferation of mesothelial cells and fibroblasts. In the presence of glycerol or amino acids synthesis of mesothelium-derived growth factors for fibroblasts is augmented. Finally, fibroblasts produce factors that inhibit the proliferation of the mesothelial cells, and this effect is potentiated in the presence of amino acids. Conclusions: Cytotoxicity of the osmotic solutes measured by the increased damage is significantly reduced during in vitro kinetic study when the concentration of these solutes is gradually lowered. Presence of osmotic solutes in the medium affects synthesis of growth factors derived from mesothelium, fibroblasts, or peritoneal leukocytes, which affect the proliferation of mesothelial cells or fibroblasts.