ORIGINAL ARTICLE Cell Research (2011): 1-13 Specific lectin biomarkers for isolation of human pluripotent stem cells identified through array-based glycomic analysisYu-Chieh Wang1,2, Masato Nakagawa3, Ibon Garitaonandia1,2, Ileana Slavin1,2, Gulsah Altun1,2, Robert M Lacharite1,2, Kristopher L Nazor1,2, Ha T Tran1,2, Candace L Lynch1,2, Trevor R Leonardo1,2, Ying Liu1,2,7, Suzanne E Peterson1,2, Louise C Laurent1,2,7, Shinya Yamanaka3,4,5,6 and Jeanne F Loring1,2 1The Scripps Research Institute, Department of Chemical Physiology, La Jolla, CA 92037, USA2The Scripps Research Institute, Center for Regenerative Medicine, 10550 N. Torrey Pines Rd., SP-3021, La Jolla, CA 92037, USA 3Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan 4Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto, Japan 5Japan Science and Technology Agency, Yamanaka iPS Cell Special Project, Kawaguchi, Japan 6Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158-2261, USA 7Department of Reproductive Medicine, University of California, San Diego, La Jolla, CA, USA
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. Cell Research advance online publication 6 September 2011; doi: 10.1038/cr.2011.148 Keywords: lectins; glycosylation; pluripotency biomarkers; human pluripotent stem cell; induced pluripotent stem cells; embryonic stem cells |
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