Yoshinao Z. Hosaka

Chondroitin sulfate/dermatan sulfate (CS/DS) is a member of glycosaminoglycans (GAGs) found in animal tissues. Major CS/DS subclasses, O, A, C, D, and E units, exist based on the sulfation pattern in d ‐glucuronic acid (GlcA) and N ‐acetyl‐ d ‐galactosamine repeating units. DS is formed when GlcA is epimerized into l ‐iduronic acid. Our study aimed to analyze the CS/DS profile in 3T3‐L1 cells before and after adipogenic induction. CS/DS contents, molecular weight (Mw), and sulfation pattern were analyzed by using high‐performance liquid chromatography. CS/DS synthesis‐ and sulfotransferase‐related genes were analyzed by reverse transcription real‐time PCR. CS/DS amount was significantly decreased in the differentiated (DI) group compared to the non‐differentiated (ND) group, along with a lower expression of CS biosynthesis‐related genes, chondroitin sulfate N ‐acetylgalactosaminyltransferase 1 and 2, as well as chondroitin polymerizing factor. GAGs in the DI group also showed lower Mw than those of ND. Furthermore, the A unit was the major CS/DS in both groups, with a proportionally higher CS‐A in the DI group. This was consistent with the expression of carbohydrate sulfotransferase 12 that encodes chondroitin 4‐ O ‐sulfotransferase, for CS‐A formation. These qualitative and quantitative changes in CS/DS and CS/DS‐synthases before and after adipocyte differentiation reveal valuable insights into adipocyte development.