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.
Horiuchi
T
T
Division of Applied Life Science, Graduate School of University of East Asia, Yamaguchi, Japan.
Ishiwata
M
M
Kumano
K
K
Sumida
Y
Y
D000418:Albumins / Q000494:pharmacology; D000818:Animals; D002455:Cell Division; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D003094:Collagen; D003470:Culture Media; D004247:DNA / Q000096:biosynthesis*; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000378:metabolism* / Q000502:physiology; D005947:Glucose / Q000494:pharmacology; D009852:Omentum / Q000033:anatomy & histology; D051381:Rats; D017208:Rats, Wistar
D000418:Albumins; D003470:Culture Media; D003094:Collagen; D004247:DNA; D005947:Glucose