Summary
The pluripotent nature of human embryonic stem cells (hESCs) is based on their potential to form every cell type in the body. Prior to use in directed differentiation strategies, these cells need to be thoroughly characterized. The large number of glycoproteins and carbohydrates that exist on the cell surface provide an excellent opportunity for characterizing hESCs and a means to delineate pluripotent and differentiated cell types. A panel of 14 lectins, based on their specificity for a variety of carbohydrates and carbohydrate linkages, along with stage-specific embryonic antigen-4 (SSEA-4), have been chosen to examine hESCs for other potential pluripotent markers. These studies have been achieved by binding quantitation by flow cytometry and binding localization in adherent colonies by immunocytochemistry. We have shown that certain lectins may be used as markers that are associated with the pluripotent state of hESCs because binding percentages and binding localization of these lectins are similar to those of SSEA-4. This presents options for systematic classification of pluripotent hESCs and for distinguishing differentiated hESC types based on glycan presentation that accompanies differentiation.
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References
Shevinsky, L.H., Knowles, B.B., Damjanov, I., and Solter, D. (1982) Monoclonal antibody to murine embryos defines a stage-specific embryonic antigen expressed on mouse embryos and human teratocarcinoma cells. Cell, 30, 697–705.
Kannagi, R., Cochran, N.A., Ishigami, F., Hakomori, S., Andrews, P.W., Knowles, B.B., and Solter, D. (1983) Stage-specific embryonic antigens (SSEA-3 and -4) are epitopes of a unique globo-series ganglioside isolated from human teratocarcinoma cells. EMBO J, 2, 2355–2361.
Draper, J.S., Pigott, C., Thomson, J.A., and Andrews, P.W. (2002) Surface antigens of human embryonic stem cells changes upon differentiation in culture. J Anat, 200, 249–258.
Andrews, P.W., Goodfellow, P.N., Shevinsky, L.H., Bronson, D.L., and Knowles, B.B. (1982) Cell-surface antigens of a clonal human embryonal carcinoma cell line: morphological and antigenic differentiation in culture. Int J Cancer, 29, 523–531.
Ruan, S., Raj, B.K., and Lloyd, K.O. (1999) Relationship of glycosyltransferases and mRNA levels to ganglioside expression in neuroblastoma and melanoma cells. J Neurochem, 72, 514–521.
Belloni, P.N., Nicolson, G.L. (1988) Differential expression of cell surface glycoproteins on various organ-derived microvascular endothelia and endothelial cell cultures. J Cell Physiol, 136, 398–410.
Takagi, Y., Talbot, N.C., Rexroad, C.E.J., and Pursel, V.G. (1997) Identification of pig primordial germ cells by immunocytochemistry and lectin binding. Mol Reprod Dev, 46, 567–580.
Brown, P.J., Stephenson, T.J. (1989) Ulex europaeus agglutinin 1 lectin histochemical staining of dog hepatocellular and bile duct carcinomas. Res Vet Sci, 46, 421–423.
Chapman, S.A., Bonshek, R.E., Stoddart, R.W., Jones, C.J., Mackenzie, K.R., O’Donoghue, E., McLeod, D. (1995) Glycoconjugates of the human trabecular meshwork: a lectin histochemical study. Histochem J, 27, 869–881.
Laitinen, L., Juusela, H., and Virtanen, I. (1990) Binding of the blood group-reactive lectins to human adult kidney specimens. Anat Rec, 226, 10–17.
Mitalipova, M., Calhoun, J., Shin, S., Wininger, D., Schulz, T., Noggle, S., Venable, A., Lyons, I., Robins, A., and Stice, S. (2003) Human embryonic stem cell lines derived from discarded embryos. Stem Cells, 21, 521–526.
Venable, A., Mitalipova, M., Lyons, I., Jones, K., Shin, S., Pierce, M., and Stice, S.L. (2005) Lectin binding profiles of SSEA-4 enriched, pluripotent human embryonic stem cell surfaces. BMC Dev Biol, 5, 15.
Acknowledgments
This work was supported in part by funding provided by the Georgia Tech/Emory Center (GTEC) for the Engineering of Living Tissues, an ERC Program of the National Science Foundation under award no. EEC- 9731643. This work was also supported in part by funding from Bresagen Inc. and from the Department of Chemical and Life Science Engineering at Virginia Commonwealth University.
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© 2007 Humana Press
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Rao, R., Johnson, A., Stice, S. (2007). Cell Surface Markers in Human Embryonic Stem Cells. In: Vemuri, M.C. (eds) Stem Cell Assays. Methods in Molecular Biology™, vol 407. Humana Press. https://doi.org/10.1007/978-1-59745-536-7_5
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DOI: https://doi.org/10.1007/978-1-59745-536-7_5
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