Glycomics of bone marrow-derived mesenchymal stem cells can be used to evaluate their cellular differentiation stage

Abstract

Human mesenchymal stem cells (MSCs) are adult multipotent progenitor cells. They hold an enormous therapeutic potential, but at the moment there is little information on the properties of MSCs, including their surface structures. In the present study, we analyzed the mesenchymal stem cell glycome by using mass spectrometric profiling as well as a panel of glycan binding proteins. Structural verifications were obtained by nuclear magnetic resonance spectroscopy, mass spectrometric fragmentation, and glycosidase digestions. The MSC glycome was compared to the glycome of corresponding osteogenically differentiated cells. More than one hundred glycan signals were detected in mesenchymal stem cells and osteoblasts differentiated from them. The glycan profiles of MSCs and osteoblasts were consistently different in biological replicates, indicating that stem cells and osteoblasts have characteristic glycosylation features. Glycosylation features associated with MSCs rather than differentiated cells included high-mannose type N-glycans, linear poly-N-acetyllactosamine chains and α2-3-sialylation.

N-Glycosylation Profile of Undifferentiated and Adipogenically Differentiated Human Bone Marrow Mesenchymal Stem Cells: Towards a Next Generation of Stem Cell Markers

“…6). The most abundant biantennary N-glycans H5N4F1 (m/z 2244.1) and S1H5N4F1 (m/z 2605.3) were found to be core-fucosylated in undifferentiated MSCs and adipogenically differentiated MSCs since their fragmentation generated the two diagnostic One signal G1H5N4 at m/z 2461.2 was found to correspond to an N-glycan, which bears N-glycolylneuraminic acid and was also observed in a previous study [37] in MSCs, but this glycan was only found as a trace. This contaminant is most likely stemming from cell culture media that contain animal-derived components [38,39].…”

Anti-Human Embryonic Stem Cell Monoclonal Antibody Hesca-2 Binds to a Glycan Epitope Commonly Found on Carcinomas

“…Other glycan markers have also been used to identify embryonic as wells as mesenchymal (adult) stem cells, such as high-mannose type N-glycans, linear poly-Nacetyllactosamine chains, alpha2-3-sialylation, and sialyl Lewis x epitopes [35][36][37]. Many of these are also cancerassociated antigens.…”

Cell Surface Sialylated N-Glycan Alterations during Development

“…High mannose-type N-glycans are characteristic for undifferentiated bone-marrow derived MSCs from humans (73,75) and equines (76). Biantennary N-glycans are detected in adipose tissue-derived hMSCs (72).…”

“…Biantennary N-glycans are detected in adipose tissue-derived hMSCs (72). Hybrid type N-glycans are found to be the less abundant type of N-glycosylation in undifferentiated and adipogenically differentiated hMSCs (73,75). In mice, an extraordinary branch is demonstrated by expression of GnT-V (the corresponding gene is Mgat5) in neural progenitor cells that have a self-renewal ability and multipotency (77).…”