Glycan Profiling Shows Unvaried N-Glycomes in MSC Clones with Distinct Differentiation Potentials

Abstract

Different cell types have different N-glycomes in mammals. This means that cellular differentiation is accompanied by changes in the N-glycan profile. Yet when the N-glycomes of cell types with differing fates diverge is unclear. We have investigated the N-glycan profiles of two different clonal populations of mesenchymal stromal cells (MSCs). One clone (Y101), when differentiated into osteoblasts, showed a marked shift in the glycan profile toward a higher abundance of complex N-glycans and more core fucosylation. Yet chemical inhibition of complex glycan formation during osteogenic differentiation did not prevent the formation of functional osteoblasts. However, the N-glycan profile of another MSC clone (Y202), which cannot differentiate into osteoblasts, was not significantly different from that of the clone that can. Interestingly, incubation of Y202 cells in osteogenic medium caused a similar reduction of oligomannose glycan content in this non-differentiating cell line. Our analysis implies that the N-glycome changes seen upon differentiation do not have direct functional links to the differentiation process. Thus N-glycans may instead be important for self-renewal rather than for cell fate determination.

Surface glycan pattern of canine, equine, and ovine bone marrow‐derived mesenchymal stem cells

“…The lectin‐binding signals indicated that the MSC surface of the three investigated species contained a complex glycan pattern with high mannose N‐linked glycans predominating over O ‐glycans, which were expressed at low levels. This result agrees with that reported in other glycomic studies on hMSCs . O ‐glycans contained the core 1 disaccharide (Galβ1,3GalNAc) (named T antigen) (Jacalin and PNA reactivity), the terminal αGalNAc and Tn antigen (the simplest mucin O ‐glycan made by a single GalNAc linked to serine or threonine) (SBA), the oligosaccharides terminating with GalNAcα1,3( l ‐Fucα1,2)Galβ1,3/4GlcNAcβ1 (DBA), and the disialyl T‐antigen sequence NeuNAcα2,3Galβ1,3(±NeuNAcα2,6)GalNAc (MAL II).…”

“…Lectins are particularly well suited for discriminating glycoconjugates because of their specificity and ability to distinguish sugar isomers, as well as branching, linkage, and terminal modifications, of complex glycans . Lectins have been used for hMSC glycomic analysis in several techniques, such as flow cytometry , lectin histochemistry , and lectin microarray . These techniques are laborious and time‐consuming, however, and the ability to measure large sample sets is limited.…”

Modeling Glycan Processing Reveals Golgi-Enzyme Homeostasis upon Trafficking Defects and Cellular Differentiation

“…They can differentiate into several cell types, and the Y101 immortalized human MSC line readily differentiates into osteoblasts under suitable growth conditions ( James et al., 2015 ). The glycan profile of Y101-derived osteoblasts is markedly different from that of the parent MSC lines ( Wilson et al., 2016 ). Glycosylation has been shown to modulate the ability of this MSC line to undergo osteogenesis ( Wilson et al., 2018 ).…”

Glycans modify mesenchymal stem cell differentiation to impact the function of resulting osteoblasts

“…Moreover, the importance of glycan processing for the development of complex life is well recognized, given many described developmental defects caused by aberrant glycosylation ( Hennet and Cabalzar, 2015 ). It is also well documented that protein-linked glycan compositions undergo large changes during differentiation events in mammals, thereby giving rise to large cell type-dependent variations in glycan profiles ( An et al, 2012 ; Hamouda et al, 2013 ; Hasehira et al, 2012 ; Wilson et al, 2016 ). However, it is largely unknown whether these glycosylation changes are functionally contributing to the differentiation process itself, potentially altering the function of the differentiated cells, or are mere bystanders of cell-specification processes.…”