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EMBRYONIC STEM CELLS: CHARACTERIZATION SERIES

The Lectin Dolichos Biflorus Agglutinin Recognizes Glycan Epitopes on the Surface of Murine Embryonic Stem Cells: A New Tool for Characterizing Pluripotent Cells and Early Differentiation

^aCenter for Complex Carbohydrate Research and Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA;
^bDepartment of Molecular Biosciences, University of Adelaide, Adelaide, Australia

Key Words. Embryonic stem cells • Primitive ectoderm • Lectin • Differentiation

Correspondence: Stephen Dalton, Ph.D., Coverdell Center for Biomedical and Health Sciences, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, 30602 Telephone: 706-583-0480; Fax: 706-583-0480; e-mail: sdalton{at}uga.edu

Received April 14, 2006; accepted for publication December 5, 2006.
First published online in STEM CELLS EXPRESS   December 14, 2006.



Cell surface markers are key tools that are frequently used to characterize and separate mixed cell populations. Existing cell surface markers used to define murine embryonic stem cells (mESCs) such as stage-specific embryonic antigen 1 (SSEA1), Forssman antigen (FA), alkaline phosphatase (AP), and CD9 are limiting, however, because they do not unambiguously define the pluripotent state and are not reliable indicators of differentiation commitment. To identify glycan cell surface markers that would circumvent this problem, we used a panel of 18 lectins to identify epitopes specifically elevated on the surface of mESCs, which, during differentiation, decrease with kinetics that precede currently used markers such as CD9, SSEA1, FA, and AP. The anticipated outcome of this analysis was to identify glycans that have utility as reliable mESC markers and high-resolution readouts for early differentiation commitment. Here, we show that the lectin Dolichos biflorus agglutinin (DBA) recognizes -N-acetylgalactosamine (GalNAc) cell surface epitopes on mESCs (CD9^high SSEA1^high AP^high DBA^high). These glycan epitopes decline markedly in cells undergoing the first definable step of differentiation, the transition from mESCs to primitive ectoderm (CD9^high SSEA1^high AP^high DBA^low). Loss of GalNAc epitopes is, therefore, the earliest cell surface change that can be assigned to differentiating cells, and the only cell surface marker known to be tightly associated with the pluripotent state. The lectin DBA is, therefore, a useful tool to characterize mESC cultures by nondestructive approaches, an indicator of differentiation commitment, and a predictor of developmental potency.

Disclosure of potential conflicts of interest is found at the end of this article.