alexa Podocalyxin-Targeting Comparative Glycan Profiling Reveals Difference between Human Embryonic Stem Cells and Embryonal Carcinoma Cells
ISSN: 2153-0637

Journal of Glycomics & Lipidomics
Open Access

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Research Article

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

Yoko Itakura1,2, Atsushi Kuno2, Masashi Toyoda1, Akihiro Umezawa3 and Jun Hirabayashi2,4*
1Vascular Medicine, Research Team for Geriatric Medicine, Tokyo Metropolitan Institute of Gerontology, 35-2 Sakae-cho, Itabashi-ku, Tokyo 173-0015, Japan
2Lectin Application and Analysis Team, Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 2, 1-1-1 Umezono, Tsukuba, Ibaraki 305-8568, Japan
3Department of Reproductive Biology, National Institute for Child Health and Development, 2-10-1 Okura, Setagaya, Tokyo 157-8535, Japan
4Glycan and Lectin Engineering Team, Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 2, 1-1-1 Umezono, Tsukuba, Ibaraki 305-8568, Japan
Corresponding Author : Jun Hirabayashi
Glycan and Lectin Engineering Team
Research Center for Stem Cell Engineering
National Institute of Advanced Industrial Science and Technology (AIST)
Tsukuba Central 2, 1-1-1 Umezono
Tsukuba, Ibaraki 305-8568, Japan
Tel: +81-29-861-3124
Fax: +81-29-861-3125
E-mail: [email protected]
Received January 19, 2013; Accepted February 05, 2013; Published February 08, 2013
Citation: Itakura Y, Kuno A, Toyoda M, Umezawa A, Hirabayashi J (2013) Podocalyxin-Targeting Comparative Glycan Profiling Reveals Difference between Human Embryonic Stem Cells and Embryonal Carcinoma Cells. J Glycomics Lipidomics S5:004. doi: 10.4172/2153-0637.S5-004
Copyright: © 2013 Itakura Y, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
 

Abstract

Background: Human embryonic stem cells (hESCs) and human embryonal carcinoma cells (hECCs) have been extensively used for stem cell research. Although these cells are known to share many properties including high developmental capability and cell surface antigens, their origins are basically different: hECSs are derived from inner cell mass of blastocysts, while hECCs are from malignant tumors. Thus, the lack of a good method to differentiate these pluripotent cells remains a critical issue to diagnose tumorigenic potential of pluripotent stem cells for their medical applications. In this context, development of specific markers to distinguish hESCs from hECCs is also of clinical value. Method: In this study, we focused our glycan analysis on a carbohydrate-rich glycoprotein, podocalyxin, known as a carrier of TRA-1-60 and TRA-1-81 antigens, which represent hESC glycan markers. The target glycoprotein semi-quantified by immunoblotting was enriched from the cell extracts by immunoprecipitation, and the glycosylation differences occurring between hESCs and hECCs were systematically analyzed by an advanced technology of lectin microarray, antibody-overlay lectin profiling (ALP). Profiles of human embryonic bodies (hEBs) differentiated from hESCs were also analyzed. Results and Conclusion: A glycan profile of podocalyxin from hECCs was significantly different from that of hESCs. Lectin signals corresponding to α2-6 linked sialic acid were elevated in the hECCs, and glycosidase digestions further revealed significant difference in the non-reducing terminal and penultimate structures. These results demonstrate that the present procedure with focus on a particular glycoprotein could enhance relatively small but significant differences between closely related cells like hESCs and hECCs at the glycome level. The present finding will be helpful to develop a diagnostic method to distinguish undifferentiated stem cells from differentiated ones used for regenerative therapy.

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