Abstract

BackgroundMesenchymal stromal cells (MSC) are components of connective tissues and, in vitro, cell entities characterized by cell adhesion and immunophenotyping, although specific markers for their identification are lacking. Currently, MSC derived from either human bone marrow (BM-MSC) or adipose tissue (Ad-MSC) are considered the main sources of MSC for cell therapy. Eph receptors and their ligands, Ephrins, are molecules involved in cell adhesion and migration in several tissues and organs. In the current study, we analyze the pattern of Eph/Ephrin expression in MSC and evaluate the effects of blockade and stimulation of these receptor/ligand pairs on their biology.MethodsEph/Ephrin expression was analyzed in both BM-MSC and Ad-MSC by qRT-PCR. Then, we supplied BM-MSC cultures with either blocking or activating compounds to evaluate their effects on MSC proliferation, survival, and cell cycle by FACS. Changes in cytoskeleton and integrin α5β1 expression were studied in stimulated BM-MSC by immunofluorescence microscopy and FACS, respectively.ResultsHigher numbers of Eph/Ephrin transcripts occurred in BM-MSC than in Ad-MSC. In addition, the blocking of Eph/Ephrin signaling correlated with decreased numbers of BM-MSC due to increased proportions of apoptotic cells in the cultures but without variations in the cycling cells. Unexpectedly, activation of Eph/Ephrin signaling by clustered Eph/Ephrin fusion proteins also resulted in increased proportions of apoptotic MSC. In this case, MSC underwent important morphological changes, associated with altered cytoskeleton and integrin α5β1 expression, which did not occur under the blocking conditions.ConclusionsTaken together, these results suggest that Eph/Ephrin activation affects cell survival through alterations in cell attachment to culture plates, affecting the biology of BM-MSC.

Highlights

  • Mesenchymal stromal cells (MSC) are components of connective tissues and, in vitro, cell entities characterized by cell adhesion and immunophenotyping, specific markers for their identification are lacking

  • We further studied the effects of blockade/stimulation of Eph/Ephrin signaling on the survival, proliferation, and differentiation, parameters studied in numerous cell types but not in Bone marrow-derived mesenchymal stromal cells (BM-MSC), observing that the blockade of Eph-Ephrin interactions produces increased proportions of apoptotic MSC without significant changes in their proliferation or cell morphology

  • Except for EphA5, EphA6, EphA8–1, EphB1, and EphrinB3, which were expressed in MSC from the two sources, BM-MSC expressed a higher number of Eph and Ephrin transcripts than those isolated from adipose tissue (Ad-MSC), especially EphA3, A7, and B2, and EphrinA1, A3, and B2 (Fig. 2)

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Summary

Introduction

Mesenchymal stromal cells (MSC) are components of connective tissues and, in vitro, cell entities characterized by cell adhesion and immunophenotyping, specific markers for their identification are lacking. MSC derived from either human bone marrow (BM-MSC) or adipose tissue (Ad-MSC) are considered the main sources of MSC for cell therapy. Eph receptors and their ligands, Ephrins, are molecules involved in cell adhesion and migration in several tissues and organs. Mesenchymal stromal cells (MSC) were described initially in the bone marrow and later in the connective tissue of almost all organs [1].

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