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A lectin-based cell microarray approach to analyze the mammalian granulosa cell surface glycosylation profile

  • Glycoarray Section
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Abstract

The high complexity of glycome, the repertoire of glycans expressed in a cell or in an organism, is difficult to analyze and the use of new technologies has accelerated the progress of glycomics analysis. In the last decade, the microarray approaches, and in particular glycan and lectin microarrays, have provided new insights into evaluation of cell glycosylation status. Here we present a cell microarray method based on cell printing on microarray slides for the analysis of the glycosylation pattern of the cell glycocalyx. In order to demonstrate the reliability of the developed method, the glycome profiles of equine native uncultured mural granulosa cells (uGCs) and in vitro cultured mural granulosa cells (cGCs) were determined and compared. The method consists in the isolation of GCs, cell printing into arrays on microarray slide, incubation with a panel of biotinylated lectins, reaction with fluorescent streptavidin and signal intensity detection by a microarray scanner. Cell microarray technology revealed that glycocalyx of both uGCs and cGCs contains N-glycans, sialic acid terminating glycans, N-acetylglucosamine and O-glycans. The comparison of uGCs and cGCs glycan signals indicated an increase in the expression of sialic acids, N-acetylglucosamine, and N-glycans in cGCs. Glycan profiles determined by cell microarray agreed with those revealed by lectin histochemistry. The described cell microarray method represents a simple and sensitive procedure to analyze cell surface glycome in mammalian cells.

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Acknowledgments

This work was supported by the Slovak Research and Development Agency under the contract No. APVV-14-0753 and by the Slovak Scientific Grant Agency VEGA project No. 2/0162/14. This document was partially funded by the University of Bari “Aldo Moro”, Italy. Dr. N.A. Martino and Dr. G. Accogli were granted by the project ONEV (Omica e Nanotecnologie applicate agli Esseri Viventi per la diagnosi di malattie) MIUR PONa3 00134–n.254/R&C 18/05/2011 (2013–2014) and Dr. N.A. Martino was granted by the project GR-2011-02351396 (Ministry of Health, Young Researchers Grant 2011/2012), “OMICS biomarkers of xenobiotic- and radiation-induced ovarian failure in farm animals - Impact on animal production and translational research for human reproductive medicine” (2015).

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Authors and Affiliations

  1. Department of Emergency and Organ Transplantation (DETO), Veterinary Clinics and Animal Productions Unit, University of Bari Aldo Moro, SP Casamassima, km 3, 70010, Valenzano, Bari, Italy

    Gianluca Accogli & Salvatore Desantis

  2. Department of Biosciences, Biotechnologies and Biopharmaceutics (DBBB), University of Bari Aldo Moro, SP Casamassima, km 3, 70010, Valenzano, Bari, Italy

    Nicola Antonio Martino & Maria Elena Dell’Aquila

  3. Experimental Zooprophylactic Institute of Puglia and Basilicata, Via Manfredonia 20, 71121, Foggia, Italy

    Nicola Antonio Martino

  4. Department of Glycobiotechnology, Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK-84538, Bratislava, Slovakia

    Peter Gemeiner & Jaroslav Katrlík

Authors
  1. Gianluca Accogli
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  2. Salvatore Desantis
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  3. Nicola Antonio Martino
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  4. Maria Elena Dell’Aquila
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  5. Peter Gemeiner
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  6. Jaroslav Katrlík
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Corresponding author

Correspondence to Jaroslav Katrlík.

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Accogli, G., Desantis, S., Martino, N.A. et al. A lectin-based cell microarray approach to analyze the mammalian granulosa cell surface glycosylation profile. Glycoconj J 33, 717–724 (2016). https://doi.org/10.1007/s10719-016-9666-2

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  • DOI: https://doi.org/10.1007/s10719-016-9666-2

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