Glycobiology Advance Access originally published online on June 29, 2006
Glycobiology 2006 16(10):981-990; doi:10.1093/glycob/cwl019
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Use of lectins for probing differentiated human embryonic stem cells for carbohydrates
2 Department of Biological Chemistry and 3 Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI 48109
1 To whom correspondence should be addressed; e-mail: igoldste{at}umich.edu
Received on April 12, 2006; revised on June 19, 2006; accepted on June 23, 2006
The carbohydrates present on the surface of differentiated human embryonic stem cells (hESCs) 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 hESC differentiation as embryoid bodies (EBs). On the basis of staining with fluorescein-labeled lectins, we have determined the presence of both terminal and internally linked 
-D-mannopyranosyl groups, poly-N-acetyllactosaminyl chains, both 2,3- and 2,6-linked N-acetylneuraminic acid (Neu5Ac), 1,6-linked L-fucosyl, and ß-D-galactosyl groups, and more specifically, the T, Tn, and sialyl-Tn antigens. However, no 1,2-linked L-fucosyl, terminal nonreducing -D-galactosyl, N-acetyl-ß-D-glucosaminyl, nor N-acetyl--D-galactosaminyl groups were found by this approach. We also established the presence of Neu5Ac2,3/2,6-Galß1,4 GlcNAc-terminated chains on the surfaces of 12-day-old EBs, as indicated by the great enhancement of staining by Erythrina cristagalli agglutinin (ECA) 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 nonspecific binding of the lectins. A comparison with undifferentiated cell staining revealed an increase in 2,3-linked Neu5Ac as well as a change to exclusively 1,6-linked L-fucose upon differentiation.
Key words: carbohydrates / differentiation / embryoid body / human embryonic stem cells / lectin
1 The authors have not provided references for the carbohydrate-binding specificity of each lectin employed in this article. Rather they have cited several monographs, which provide the information on the carbohydrate-binding properties of the lectins employed in this study. When appropriate, citations that provide more recent or relative information are noted.
2 All carbohydrates reported in this article are of the D-configuration except for fucose, which is of the L-configuration.
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