Comparative Analysis of Proteoglycans Synthesized by Chick Corneal Stromal Cells in Cell Culture and Organ Culture

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Proteoglycans (PCs) synthesized by chick corneal stromal cells in cell culture and organ culture were metabolically radiolabelled with [^<35>S]sulfate (for glycosaminoglycans) and [^3H]leucine (for core proteins). Media, cell extracts and organ extracts from cultures were chromatographed on DEAE-Sephacel columns and separated into three fractions: the pass-through fraction (Fraction 1: nonsulfated PCs, hardly sulfated PCs, or glycoproteins with some oligosaccharides), the fraction eluted with a low salt concentration (Fraction 2: undersulfated PGs), and the fraction eluted with a high salt concentration (Fraction 3: highly sulfated PGs). The PG compositions of each fraction of cell culture and organ culture were then compared. While the proportions of highly sulfated KSPG in Fractions 3 of medium and cell extract of cell culture were both very low compared with those of medium and organ extract of organ culture, respectively, the proportions of highly sulfated CS/DS PG in Fractions 3 of those of cell culture were higher than those of organ culture. On the other hand, the proportions in the^<35>S activities of nonsulfated or undersulfated KSPG in Fractions 1 and 2 of medium and cell extract of cell culture were comparable to those of organ culture. Furthermore, the proportions of core proteins of undersulfated KSPG in Fractions 2 were higher in cell culture than in organ culture. These results show that, when the cells are cell-cultured, the degree of sulfation of KS chains decreases markedly, but the syntheses of the glycosaminoglycan backbone and core protein are maintained.
社団法人日本薬学会の論文
2001-01-01

Comparative Analysis of Proteoglycans Synthesized by Chick Corneal Stromal Cells in Cell Culture and Organ Culture

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