Specific lectin biomarkers for isolation of human pluripotent stem cells identified through array-based glycomic analysis

Abstract

Rapid and dependable methods for isolating human pluripotent stem cell (hPSC) populations are urgently needed for quality control in basic research and in cell-based therapy applications. Using lectin arrays, we analyzed glycoproteins extracted from 26 hPSC samples and 22 differentiated cell samples, and identified a small group of lectins with distinctive binding signatures that were sufficient to distinguish hPSCs from a variety of non-pluripotent cell types. These specific biomarkers were shared by all the 12 human embryonic stem cell and the 14 human induced pluripotent stem cell samples examined, regardless of the laboratory of origin, the culture conditions, the somatic cell type reprogrammed, or the reprogramming method used. We demonstrated a practical application of specific lectin binding by detecting hPSCs within a differentiated cell population with lectin-mediated staining followed by fluorescence microscopy and flow cytometry, and by enriching and purging viable hPSCs from mixed cell populations using lectin-mediated cell separation. Global gene expression analysis showed pluripotency-associated differential expression of specific fucosyltransferases and sialyltransferases, which may underlie these differences in protein glycosylation and lectin binding. Taken together, our results show that protein glycosylation differs considerably between pluripotent and non-pluripotent cells, and demonstrate that lectins may be used as biomarkers to monitor pluripotency in stem cell populations and for removal of viable hPSCs from mixed cell populations.

Structural and Quantitative Evidence for Dynamic Glycome Shift on Production of Induced Pluripotent Stem Cells

“…Thus, the development of novel cell surface markers to evaluate the properties of iPSCs and ESCs is keenly required. Toward this goal, a glycomic approach has been made by several groups (17)(18)(19)(20). In our previous study using an advanced lectin microarray technique (21), thirty-eight lectins capable of discriminating between iPSCs and SCs were statistically selected, and the characteristic features of the pluripotent state were obtained.…”

Four new induced pluripotent stem cell lines produced from northern white rhinoceros with non-integrating reprogramming factors

“…Immunocytochemistry (ICC) showed that the iPSCs were positive for canonical markers of pluripotency [OCT4 (also known as POU5F1), SOX2 and NANOG] (Figure 1b) and the iPSCs differentiated via embryoid body (EB) formation expressed markers for each of the germ layers (Figure 1c) 1,3 . qPCR analyses of the iPSCs confirmed expression of pluripotency markers (Figure 1d).…”

Podocalyxin-Targeting Comparative Glycan Profiling Reveals Difference between Human Embryonic Stem Cells and Embryonal Carcinoma Cells

“…In the authors' laboratory, as a lectin-based glycan profiling method, a unique lectin microarray system based on an evanescentfield activated fluorescence detection principle was developed [41,42]. With this system, we and other groups successfully profiled distinct sets of glycomes of both somatic and pluripotent stem cells [43][44][45][46][47].…”