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  • Interaction between Galectin-3 and Integrins Mediates Cell-Matrix Adhesion in Endothelial Cells and Mesenchymal Stem Cells.

Interaction between Galectin-3 and Integrins Mediates Cell-Matrix Adhesion in Endothelial Cells and Mesenchymal Stem Cells.

International journal of molecular sciences (2021-06-03)
Antonín Sedlář, Martina Trávníčková, Pavla Bojarová, Miluše Vlachová, Kristýna Slámová, Vladimír Křen, Lucie Bačáková
ABSTRACT

Galectin-3 (Gal-3) is a β-galactoside-binding protein that influences various cell functions, including cell adhesion. We focused on the role of Gal-3 as an extracellular ligand mediating cell-matrix adhesion. We used human adipose tissue-derived stem cells and human umbilical vein endothelial cells that are promising for vascular tissue engineering. We found that these cells naturally contained Gal-3 on their surface and inside the cells. Moreover, they were able to associate with exogenous Gal-3 added to the culture medium. This association was reduced with a β-galactoside LacdiNAc (GalNAcβ1,4GlcNAc), a selective ligand of Gal-3, which binds to the carbohydrate recognition domain (CRD) in the Gal-3 molecule. This ligand was also able to detach Gal-3 newly associated with cells but not Gal-3 naturally present on cells. In addition, Gal-3 preadsorbed on plastic surfaces acted as an adhesion ligand for both cell types, and the cell adhesion was resistant to blocking with LacdiNAc. This result suggests that the adhesion was mediated by a binding site different from the CRD. The blocking of integrin adhesion receptors on cells with specific antibodies revealed that the cell adhesion to the preadsorbed Gal-3 was mediated, at least partially, by β1 and αV integrins-namely α5β1, αVβ3, and αVβ1 integrins.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Gly-Arg-Ala-Asp-Ser-Pro (GRADSP)
Sigma-Aldrich
Anti-Integrin αV Antibody, extracellular, clone AV1, azide free, culture supernatant, clone AV1, Chemicon®
Sigma-Aldrich
Anti-Integrin α2β1 Antibody, clone BHA2.1, clone BHA2.1, Chemicon®, from mouse
Sigma-Aldrich
Mouse IgG1 Negative Control, clone Ci4, Mouse IgG1 Negative Control Monoclonal Antibody validated for use in Flow Cytometry & Immunofluorescence.
Sigma-Aldrich
Anti-Integrin β1 Antibody, clone 6S6, clone 6S6, Chemicon®, from mouse
Sigma-Aldrich
Anti-Galectin 3 antibody produced in rabbit, affinity isolated antibody
Sigma-Aldrich
Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP)
Sigma-Aldrich
Monoclonal Anti-Vinculin antibody produced in mouse, clone hVIN-1, ascites fluid
Sigma-Aldrich
Anti-Integrin β3 Antibody, clone B3A, azide free, clone B3A, Chemicon®, from mouse
Sigma-Aldrich
Anti-Integrin α3 Antibody, clone P1B5, azide free, clone P1B5, Chemicon®, from mouse
Sigma-Aldrich
Anti-Integrin αVβ3 Antibody, clone LM609, clone LM609, Chemicon®, from mouse