Japan Chemical Innovation Institute, 2-11-9 Iwamotocho, Chiyoda-ku, Tokyo 101-0032, Japan.
J Biosci Bioeng. 2012 Nov;114(5):552-5. doi: 10.1016/j.jbiosc.2012.06.007. Epub 2012 Jul 12.
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 β-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.
采用糖基供体法可以评估细胞在糖生物合成方面的功能。本研究表明，当将 12-叠氮十二烷基-β-乳糖苷（Lac-12N（3）供体）掺入 Madin-Darby 犬肾（MDCK）细胞中，并生成 GM3、GD3、唾液酸神经节苷脂（SPG）和 NeuAc-Gal-GlcNAc-Gal-GlcNAc-Lac 型寡糖时，MDCK 细胞的特征会随传代数的增加而发生变化。通过测量相对早期到晚期传代数的糖基化产物的量，结果表明存在一个优化寡糖生成的合适传代数，而较高的传代数会导致寡糖生成减少。此外，结果表明，除了唾液酸转移酶之外，控制糖产量的几种酶的活性可能会受到生物衰老的影响。
