Synthesis and secretion of glycosaminoglycans in cultured retinal pigment epithelium.

摘要: The synthesis and secretion of glycosaminoglycans (GAGs) in primary cultures feline retinal pigment epithelium (RPE) was measured. After 14 days culture, the cells were incubated for 3 with media containing 20 microCi/ml 35SO4 10 [3H]-glucosamine. GAGs precipitated from cell layer cetylpyridinium chloride ethanol, separated 0.1 M Ba acetate by cellulose electrophoresis, visualized Alcian blue staining fluorography, quantitated scanning densitometry liquid scintillation counting. predominant radioactively-labeled GAG secreted into RPE chondroitin sulfate (CS), which accounted 63% 35SO4, 54% 3H incorporated. Radioactively-labeled heparan (HS), dermatan (DS), hyaluronic acid (HA) also identified media. Heparan 24% 22% 3H, DS contained 13% 7% HA 17% incorporated GAGs. In contrast, fibroblast primarily HA, 84% profile retained different that associated HS. This 52% Dermatan 23% while CS 18% remaining HA. Total profiles determined staining, compared those freshly isolated samples. similar cultured freshly-isolated samples; however, percentage HS samples about twice as high samples, proportion higher than study demonstrates is distinct layer. addition, total similar, but not identical, to cells.