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Glycobiology Advance Access published online on June 29, 2006
Glycobiology, doi:10.1093/glycob/cwl019
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© The Author 2006. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Article

Use of Lectins for Probing Differentiated Human Embryonic Stem Cells for Carbohydrates

Kimberly A. Wearne 1, Harry C. Winter 1, Katherine O’Shea 2, and Irwin J. Goldstein 1 *

1 Dept. of Biological Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA
2 Dept. of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI, 48109, USA

* To whom correspondence should be addressed.
Irwin J. Goldstein, E-mail: igoldste{at}umich.edu


  Abstract

The carbohydrates present on the surface of differentiated human embryonic stem cells are not yet well established. Here we have employed a panel of lectins and several anti-carbohydrate antibodies to determine the carbohydrates that are present at day 12 of human embryonic stem cell differentiation as embryoid bodies. On the basis of staining with fluorescein-labeled lectins, we have determined the presence of both terminal and internally linked {alpha}-D-mannopyranosyl groups, poly N-acetyllactosaminyl chains, both {alpha}2,3- and {alpha}2,6-linked N-acetylneuraminic acid, {alpha}1,6-linked L-fucosyl, and {beta}-D-galactosyl groups, and more specifically, the T, Tn, and sialyl Tn antigens. However, no {alpha}1,2-linked L-fucosyl, terminal nonreducing {alpha}-D-galactosyl, N-acetyl-{beta}-D-glucosaminyl, nor N-acetyl-{alpha}-D-galactosaminyl groups were found by this approach. We also established the presence of Neu5Ac {alpha}2,3/2,6-Gal {beta}1,4 GlcNAc-terminated chains on the surfaces of 12-day-old embryoid bodies as indicated by the great enhancement of staining by Erythrina cristagalli agglutinin after treatment with neuraminidase. In each case, inhibition of binding by a haptenic sugar or treatment with neuraminidase was used to eliminate the possibility of non-specific binding of the lectins. A comparison with undifferentiated cell staining revealed an increase in {alpha}2,3-linked N-acetylneuraminic acid, as well as a change to exclusively {alpha}1,6-linked L-fucose upon differentiation.

Keywords: lectin/differentiation/human embryonic stem cells/embryoid body/carbohydrates.
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