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Cells, tissues, organs2015; 200(3-4); 212-226; doi: 10.1159/000433420

Cell Surface Glycan Changes in the Spontaneous Epithelial-Mesenchymal Transition of Equine Amniotic Multipotent Progenitor Cells.

Abstract: Amniotic epithelial cells (AECs) spontaneously transform into amniotic mesenchymal cells (AMCs) in vitro during cell culture. Glycocalyx was analyzed to identify the glycan pattern in AECs, AMCs and epithelial-mesenchymal transdifferentiated cells (EMTCs). Pure cell cultures were derived using cloned AEC and AMC cell lines obtained by the dilution technique from amniotic membranes. Mesenchymal cells generated by differentiation of clonal epithelial cells were considered transdifferentiated. Immunocytoscreen, in vitro multipotent differentiation and molecular characterization of EMTCs were performed. In combination with saponification and sialidase digestion, a panel of 12 lectins was used to analyze the glycan pattern of AEC, AMC and EMTC glycocalyx. Cytokeratin cell markers were lost in EMTCs and typical mesenchymal markers, such as vimentin, appeared. These cells retained their differentiation potential. Lectin histochemistry revealed a cell-specific glycan profile. Galactose (Gal)β1,4GlcNAc, Neu5Acα2,6Gal/GalNAc and N-acetyl neuraminic (sialic) acid (NeuNAc)α2,3Galβ1,3(±NeuNAcα2,6)GalNAc were highly expressed on the surface of all the amniotic cell cultures. AECs expressed asialoglycans with terminal GalNAc and GlcNAc. More highly mannosylated N-linked glycans and NeuNAcα2,3Galβ1,3GalNAc in O-linked glycans were expressed by EMTCs, but these cells had fewer glycans ending with fucose (Fuc), Gal, GlcNAc and GalNAc than AECs. GlcNAc- and GalNAc-terminating glycans were similarly expressed on the glycocalyx of the mesenchymal cell populations (EMTCs and AMCs). These results demonstrate for the first time that the spontaneous epithelial-mesenchymal transition (EMT) of equine amnion cells is characterized by cell surface glycan remodeling and that glycosylation changes result in a cell type-specific glycan profile. The glycopattern of equine amnion spontaneous EMTCs differs from EMT of tumoral cells.
Publication Date: 2015-08-26 PubMed ID: 26337136DOI: 10.1159/000433420Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

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This research study explored how the cell surfaces of certain progenitor cells, specifically equine amniotic epithelial and mesenchymal cells, change as the cells naturally transition from one type to another. The researchers analyzed cell cultures and discovered specific patterns in the glycans, a type of molecule on the cells’ surfaces, that were characteristic of each cell type and its transitional state.

Cellular Transformation and Source

In the initial phase of the study, the researchers focused on amniotic epithelial cells (AECs) as their subject of study. These cells are known to naturally transform into amniotic mesenchymal cells (AMCs) under in vitro conditions. The cells were sourced from amniotic membranes and pure cell cultures derived using clone AEC and AMC cell lines obtained by the dilution technique.

Transdifferentiation and Analysis

Transdifferentiation, in this context, refers to the process where cells originating from the epithelial line are converted into mesenchymal cells. All cells generated this way were considered transdifferentiated. Afterward, the glycan pattern in AECs, AMCs, and epithelial-mesenchymal transdifferentiated cells (EMTCs) was assessed using a technique involving 12 varieties of lectins (proteins that bind to specific sugar molecules).

Cell Marker Shift

The resulting data showed a shift in protein markers, with cytokeratin markers disappearing in the EMTCs, and another marker, vimentin, emerging. This shift confirms the transformation of the cells.

Glycan Profile

Further analysis revealed that the cells exhibited a unique glycan profile after transition. The dominant glycan molecules on all three types of amnion cells were galactose, N-acetyl neuraminic acid, and others. Interestingly, fucose, galactose, and other glycans were found to be less common on EMTCs than on AECs.

Significance of Research Findings

The findings suggest that as the cells transition from epithelial to mesenchymal cells, the remodeling of the cell surface glycans creates cell-specific glycan profiles. This discovery presents a first-time observation of cell surface glycan alterations during the spontaneous epithelial-mesenchymal transition of equine amnion cells. The results also highlight a key difference between the glycosylation patterns of spontaneous EMTCs and those of tumoral cells undergoing the same transition.

Cite This Article

APA
Lange-Consiglio A, Accogli G, Cremonesi F, Desantis S. (2015). Cell Surface Glycan Changes in the Spontaneous Epithelial-Mesenchymal Transition of Equine Amniotic Multipotent Progenitor Cells. Cells Tissues Organs, 200(3-4), 212-226. https://doi.org/10.1159/000433420

Publication

ISSN: 1422-6421
NlmUniqueID: 100883360
Country: Switzerland
Language: English
Volume: 200
Issue: 3-4
Pages: 212-226

Researcher Affiliations

Lange-Consiglio, Anna
  • Reproduction Unit, Large Animal Hospital, Universitx00E0; degli Studi di Milano, Lodi, Italy.
Accogli, Gianluca
    Cremonesi, Fausto
      Desantis, Salvatore

        MeSH Terms

        • Adipogenesis
        • Amnion / cytology
        • Animals
        • Cell Membrane / metabolism
        • Cell Shape
        • Cell Transdifferentiation
        • Cells, Cultured
        • Chondrogenesis
        • Epithelial Cells / cytology
        • Epithelial-Mesenchymal Transition
        • Female
        • Glycosylation
        • Horses
        • Immunoassay
        • Lectins
        • Mesoderm / cytology
        • Multipotent Stem Cells / cytology
        • Neurogenesis
        • Osteogenesis
        • Polysaccharides / metabolism
        • Real-Time Polymerase Chain Reaction

        Citations

        This article has been cited 6 times.
        1. Russo V, El Khatib M, di Marcantonio L, Ancora M, Wyrwa R, Mauro A, Walter T, Weisser J, Citeroni MR, Lazzaro F, Di Federico M, Berardinelli P, Cammà C, Schnabelrauch M, Barboni B. Tendon Biomimetic Electrospun PLGA Fleeces Induce an Early Epithelial-Mesenchymal Transition and Tenogenic Differentiation on Amniotic Epithelial Stem Cells. Cells 2020 Jan 27;9(2).
          doi: 10.3390/cells9020303pubmed: 32012741google scholar: lookup
        2. Buffone A, Weaver VM. Don't sugarcoat it: How glycocalyx composition influences cancer progression. J Cell Biol 2020 Jan 6;219(1).
          doi: 10.1083/jcb.201910070pubmed: 31874115google scholar: lookup
        3. Mauro A, Sanyal H, Canciello A, Berardinelli P, Russo V, Bernabò N, Valbonetti L, Barboni B. In Vitro Effect of Estradiol and Progesterone on Ovine Amniotic Epithelial Cells. Stem Cells Int 2019;2019:8034578.
          doi: 10.1155/2019/8034578pubmed: 31049069google scholar: lookup
        4. Canciello A, Russo V, Berardinelli P, Bernabò N, Muttini A, Mattioli M, Barboni B. Progesterone prevents epithelial-mesenchymal transition of ovine amniotic epithelial cells and enhances their immunomodulatory properties. Sci Rep 2017 Jun 19;7(1):3761.
          doi: 10.1038/s41598-017-03908-1pubmed: 28630448google scholar: lookup
        5. Accogli G, Desantis S, Martino NA, Dell'Aquila ME, Gemeiner P, Katrlík J. A lectin-based cell microarray approach to analyze the mammalian granulosa cell surface glycosylation profile. Glycoconj J 2016 Oct;33(5):717-24.
          doi: 10.1007/s10719-016-9666-2pubmed: 27085877google scholar: lookup
        6. Gaspari G, Lange-Consiglio A, Cremonesi F, Desantis S. Role of Glycans in Equine Endometrial Cell Uptake of Extracellular Vesicles Derived from Amniotic Mesenchymal Stromal Cells. Int J Mol Sci 2025 Feb 19;26(4).
          doi: 10.3390/ijms26041784pubmed: 40004247google scholar: lookup