Disaccharide analysis of dermal fibroblast-derived glycosaminoglycans in the three-dimensional culture.
We performed the quantitative and qualitative analysis on the main disaccharide units of glycosaminoglycans produced by human dermal fibroblasts in the 3-dimensional culture supplemented with L-ascorbic acid 2-phosphate (Asc 2-p) comparing with the monolayer culture system. The addition of Asc 2-p rendered fibroblasts to the organization of the dermis-like 3-dimensional structure in vitro without any pre-treatments with the plastic dish. Main disaccharide units were analyzed using HPLC after 1-phenyl-3-methyl-5-pyrasolone (PMP) labeling. The addition of Asc 2-p significantly increased the total amount of main disaccharide units and, furthermore, the composition revealed it to be more similar to that of the dermis. This 3-dimensional culture may offer a simple and useful system to investigate the glycosaminoglycan metabolism of human dermal fibroblasts in vitro.
Ishikawa
O
O
Department of Dermatology, Gunma University School of Medicine, Japan.
Yokoyama
Y
Y
Miyachi
Y
Y
D001205:Ascorbic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004187:Disaccharides / Q000032:analysis* / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000032:analysis* / Q000737:chemistry*; D006652:Histological Techniques; D006801:Humans; D012867:Skin / Q000166:cytology* / Q000378:metabolism
D004187:Disaccharides; D006025:Glycosaminoglycans; C011669:ascorbate-2-phosphate; D001205:Ascorbic Acid