RESUMEN
The cell function on saccharide biosynthesis can be evaluated by employing the saccharide primer method. This study demonstrated that the characteristics of Madin-Darby canine kidney (MDCK) cells changed in relation with passage number when 12-azidododecyl ß-lactoside (Lac-12N(3) primer) was incorporated into MDCK cells and afforded GM3-, GD3-, sialylparagloboside (SPG), and NeuAc-Gal-GlcNAc-Gal-GlcNAc-Lac-type oligosaccharides. By measuring the amount of glycosylated products from relatively early to late passage numbers, results showed that there was an appropriate passage number that optimized oligosaccharide production and that the higher passage number resulted to a decrease in oligosaccharide production. Moreover, results suggested that aside from sialyltransferase, the activity of several kinds of enzymes that control the amount of saccharide production was presumably affected depending upon the biological senescence.
Asunto(s)
Azidas/metabolismo , Lactosa/análogos & derivados , Oligosacáridos/biosíntesis , Animales , Azidas/química , Técnicas de Cultivo de Célula , Perros , Glicósidos/metabolismo , Glicosilación , Lactosa/química , Lactosa/metabolismo , Células de Riñón Canino Madin Darby , Oligosacáridos/química , Oligosacáridos/aislamiento & purificación , Sialiltransferasas/análisisRESUMEN
The efficient production of ganglioside analogues was accomplished using RERF-LC-AI cells cultured in HYPERFlask (High Yield PERformance Flask). Eight kinds of ganglioside analogues (GM3, GM2, sialylparagloboside, GD3, di-sialylated lacto-N-tetraose, and another three kinds of analogues with intricate structures) were synthesized by the saccharide primer method using lung squamous-cell carcinoma line RERF-LC-AI and 12-azidododecyl ß-lactoside primer. The yield for each analogue obtained using HYPERFlask was higher than yields obtained from 100-mm dishes.