DNA synthesis of cultured mesothelial cells in a high permeable state

To understand the mechanism of regeneration of the mesothelial cell in a high permeable state, in vitro experiments with cultured mesothelial cells were carried out using a type I collagen-coated multititer plate. Prior to cell seeding, each plate was filled with 100 microL of glucose solution at concentrations of 30 mmol/L, 90 mmol/L, and 150 mmol/L, and minimum essential medium (MEM), respectively. After immersion for one to five weeks at 37 degrees C, mesothelial cells were seeded at a density of 3 x 10(4)/cm2, then glucose was added at varying final concentrations between 0 and 150 mmol/L and/or albumin was added at a final concentration of 2 g/dL. After days 2 and 4, DNA synthesis was measured by incorporation of 5-bromo-2'-deoxy-uridine (BrdU). There was no significant difference of DNA synthesis between groups with and without one-week glucose immersion when mesothelial cells were cultured with only 10% FCS/MEM. However, suppression of DNA synthesis at 2 g/dL of albumin was seen no matter what concentration of glucose supplement was used. Without a high concentration of albumin, DNA synthesis was dose-dependent on glucose. The longer the immersion period, the lesser the suppression. It is speculated that process of glycation with collagen may relate to growth of mesothelial cells.