HS purified from OCM (A) and SCM (B) was digested with heparinase enzymes to generate HS disaccharides, which were fluorescently labelled with BODIPY and separated by SAX-HPLC over a 45 minute, 0-1.5 M NaCl gradient, (A and B, black lines). HS disaccharide standards were separated over the same gradient as a reference guide (A and B, dashed lines). Relative abundances of the eight HS standard disaccharides in each conditioned medium sample were calculated as a percentage of total HS (C). A summary table detailing the composition of OCM HS and SCM HS is also presented (D). SCM HS was found to be more highly sulfated than OCM HS, with an average of 1.02 sulfates per disaccharide compared to 0.75 sulfates per disaccharide in OCM HS (D). SCM HS contained a higher proportion of di- and tri-sulfated disaccharides (disaccharides 4, 5, 8 and 6) and the singly 6-O-sulfated disaccharide (disaccharide 2) compared to OCM HS (C), which contained a higher proportion of the unsulfated disaccharide (disaccharide 1) (C, D).
(UA) uronic acid, (GlcN) glucosamine, (NAc) N-acetyl, (NS) N-sulfate, (6OS) 6-O-sulfate, (2OS) 2-O-sulfate.