Vessel Wall-Derived Mesenchymal Stromal Cells Share Similar Differentiation Potential and Immunomodulatory Properties with Bone Marrow-Derived Stromal Cells

Abstract

Purpose. This study is aimed at investigating the phenotype, differentiation potential, immunomodulatory properties, and responsiveness of saphenous vein vessel wall-derived mesenchymal stromal cells (SV-MSCs) to various TLR ligands and proinflammatory cytokines, as well as comparing their features to those of their bone marrow-derived counterparts (BM-MSCs). Methods. SV-MSCs were isolated by enzymatic digestion of the saphenous vein vessel wall. Phenotype analysis was carried out by flow cytometry and microscopy, whereas adipogenic, chondrogenic, and osteogenic differentiation potentials were tested in in vitro assays. For comparative analysis, the expression of different stemness, proliferation, and differentiation-related genes was determined by Affymetrix gene array. To compare the immunomodulatory properties of SV-MSCs and BM-MSCs, mixed lymphocyte reaction was applied. To investigate their responses to various activating stimuli, MSCs were treated with TLR ligands (LPS, PolyI:C) or proinflammatory cytokines (TNFα, IL-1β, IFNγ), and the expression of various early innate immune response-related genes was assessed by qPCR, while secretion of selected cytokines and chemokines was measured by ELISA. Results. The isolated SV-MSCs were able to differentiate into bone, fat, and cartilage cells/direction in vitro. SV-MSCs expressed the most important MSC markers (CD29, CD44, CD73, CD90, and CD105) and shared almost identical phenotypic characteristics with BM-MSCs. Their gene expression pattern and activation pathways were close to those of BM-MSCs. SV-MSCs showed better immunosuppressive activity inhibiting phytohemagglutinin-induced T lymphocyte proliferation in vitro than BM-MSCs. Cellular responses to treatments mimicking inflammatory conditions were comparable in the bone marrow- and saphenous vein-derived MSCs. Namely, similar to BM-MSCs, SV-MSCs secreted increased amount of IL-6 and IL-8 after 12- or 24-hour treatment with LPS, PolyI:C, TNFα, or IL-1β, compared to untreated controls. Interestingly, a different CXCL-10/IP-10 secretion pattern could be observed under inflammatory conditions in the two types of MSCs. Conclusion. Based on our results, cells isolated from saphenous vein vessel wall fulfilled the ISCT’s (International Society for Cellular Therapy) criteria for multipotent mesenchymal stromal cells, and no significant differences in the phenotype, gene expression pattern, and responsiveness to inflammatory stimuli could be observed between BM-MSCs and SV-MSCs, while the latter cells have more potent immunosuppressive activity in vitro. Further functional assays have to be performed to reveal whether SV-MSCs could be useful for certain regenerative therapeutic applications or tissue engineering purposes.

Licensing effects of inflammatory factors and TLR ligands on the regenerative capacity of adipose-derived mesenchymal stem cells

“…The immunomodulatory effectiveness of MSCs is contingent upon the nature and intensity of inflammatory signals received, such as interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and Toll-like receptor (TLR)-mediated activation. Under specific inflammatory stimuli (IFN-γ, TNF-α, TLR-mediated activation), MSCs transform, becoming antiapoptotic, proangiogenic, and immunosuppressive ( Meisel et al, 2004 ; Krampera et al, 2006 ; Ghannam et al, 2010 ; Saparov et al, 2016 ; Vereb et al, 2020 ). They contribute to inflammation reduction through the secretion of factors like interleukin-6 (IL-6), indoleamine 2,3-dioxygenase (IDO), HLA G5 (human leukocyte antigen G5), interleukin-10 (IL-10), transforming growth factor beta-1 (TGFb1), hepatocyte growth factor (HGF), HOX-1, IL-1Ra (IL-1 receptor antagonist), prostaglandin E2 (PGE2), and through cell-cell contact ( Bartholomew et al, 2002 ; Meisel et al, 2004 ; Aggarwal SP and Pittenger, 2005 ; Saparov et al, 2016 ).…”

“…The immunomodulatory potential of MSCs thus hinges on their response to specific inflammatory cues ( Renner et al, 2009 ; Li et al, 2012 ). Mesenchymal stem cells (MSCs) actively produce a diverse array of chemokines and adhesion molecules, including ligands for CXC chemokine receptor 3 (CXCR3), C-C chemokine receptor type 5 (CCR5), intercellular adhesion molecule 1 (ICAM-1/CD54), and vascular cell adhesion molecule 1 (VCAM-1) ( Vereb et al, 2016 ; Vereb et al, 2020 ). The pronounced expression of CXCR3 in effector and memory T cells underscores the pivotal significance of MSC-generated chemokines, particularly CXCL9 (chemokine ligand 9), CXCL10 (chemokine ligand 10), and CXCL11 (chemokine ligand 11).…”

“…TLR receptors are one of the most ancient components of defense against pathogens and innate immunity. Tests with LPS (TLR4) and PolyI:C (TLR3) showed that MSC lifted their IL-6 and IL-8 expression ( Fuenzalida et al, 2016 ; Park et al, 2016 ; Vereb et al, 2016 ; Vereb et al, 2020 ; Szucs et al, 2023 ), and the TLR3 manifest a more potent immunosuppressive phenotype ( Fuenzalida et al, 2016 ; Park et al, 2016 ).…”

The Characteristics and Survival Potential Under Sub-lethal Stress of Mesenchymal Stromal/Stem Cells Isolated from the Human Vascular Wall

“…A comparison performed between human saphenous vein (SV-MSCs) and bone marrow MSCs (BM-MSCs) elucidated that these 2 cell populations share phenotype, gene signature and differentiation potential, whereas the SV-MSCs resulted more responsive to inflammatory stimulation by exerting a higher immunosuppressive activity on human peripheral blood mononuclear cells (PBMCs) proliferation in vitro. 34 …”

Bone marrow mesenchymal stem cells enhance angiogenesis and promote fat retention in fat grafting via polarized macrophages

“…MSCs are the most widely used stem cells. Compared with other types of stem cells, BM-MSCs have significant immunomodulatory abilities [ 14 ]. The interaction between immune cells and stem cells in tissue regeneration has been known for a long time.…”