Intact cell adhesion to glycan microarrays

Leonardo Nimrichter, Ari Gargir, Monica Gortler, Rom T. Altstock, Avi Shtevi, Oori Weisshaus, Ella Fire, Nir Dotan, Ronald Lee Schnaar

Research output: Contribution to journalArticle

Abstract

A rapid and reproducible method was developed to detect and quantify carbohydrate-mediated cell adhesion to glycans arrayed on glass slides. Monosaccharides and oligosaccharides were covalently attached to glass slides in 1.7-mm-diameter spots (200 spots/slide) separated by a Teflon gasket. Primary chicken hepatocytes, which constitutively express a C-type lectin that binds to nonreducing terminal N-acetylglucosamine residues, were labeled with a fluorescent dye and incubated in 1.3-μL aliquots on the glycosylated spots. After incubating to allow cell adhesion, nonadherent cells were removed by immersing the slide in phosphate buffered saline, inverting, and centrifuging in a sealed custom acrylic chamber so that cells on the derivatized spots were subjected to a uniform and controlled centrifugal detachment force while avoiding an air-liquid interface. After centrifugation, adherent cells were fixed in place and detected by fluorescent imaging. Chicken hepatocytes bound to nonreducing terminal GlcNAc residues in different linkages and orientations but not to nonreducing terminal galactose or N-acetylgalactosamine residues. Addition of soluble GlcNAc (but not Gal) prior to incubation reduced cell adhesion to background levels. Extension of the method to CD4+ human T-cells on a 45-glycan diversity array revealed specific adhesion to the sialyl Lewis x structure. The described method is a robust approach to quantify selective cell adhesion using a wide variety of glycans and may contribute to the repertoire of tools for the study of glycomics.

Original languageEnglish (US)
Pages (from-to)197-203
Number of pages7
JournalGlycobiology
Volume14
Issue number2
DOIs
StatePublished - Feb 2004

Fingerprint

Cell adhesion
Microarrays
Cell Adhesion
Polysaccharides
Glass
Hepatocytes
Chickens
Glycomics
C-Type Lectins
Acetylgalactosamine
T-cells
Acetylglucosamine
Centrifugation
Monosaccharides
Polytetrafluoroethylene
Oligosaccharides
Galactose
Fluorescent Dyes
Acrylics
Adhesion

Keywords

  • CD4
  • Glycomics
  • Hepatocyte
  • Lectins
  • Oligosaccharides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Nimrichter, L., Gargir, A., Gortler, M., Altstock, R. T., Shtevi, A., Weisshaus, O., ... Schnaar, R. L. (2004). Intact cell adhesion to glycan microarrays. Glycobiology, 14(2), 197-203. https://doi.org/10.1093/glycob/cwh022

Intact cell adhesion to glycan microarrays. / Nimrichter, Leonardo; Gargir, Ari; Gortler, Monica; Altstock, Rom T.; Shtevi, Avi; Weisshaus, Oori; Fire, Ella; Dotan, Nir; Schnaar, Ronald Lee.

In: Glycobiology, Vol. 14, No. 2, 02.2004, p. 197-203.

Research output: Contribution to journalArticle

Nimrichter, L, Gargir, A, Gortler, M, Altstock, RT, Shtevi, A, Weisshaus, O, Fire, E, Dotan, N & Schnaar, RL 2004, 'Intact cell adhesion to glycan microarrays', Glycobiology, vol. 14, no. 2, pp. 197-203. https://doi.org/10.1093/glycob/cwh022
Nimrichter L, Gargir A, Gortler M, Altstock RT, Shtevi A, Weisshaus O et al. Intact cell adhesion to glycan microarrays. Glycobiology. 2004 Feb;14(2):197-203. https://doi.org/10.1093/glycob/cwh022
Nimrichter, Leonardo ; Gargir, Ari ; Gortler, Monica ; Altstock, Rom T. ; Shtevi, Avi ; Weisshaus, Oori ; Fire, Ella ; Dotan, Nir ; Schnaar, Ronald Lee. / Intact cell adhesion to glycan microarrays. In: Glycobiology. 2004 ; Vol. 14, No. 2. pp. 197-203.
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