MSCs’ conditioned media cytokine and growth factor profiles and their impact on macrophage polarization

Abstract

Background. There is a growing body of evidence that multipotent mesenchymal stromal cells’ (MSCs’) remarkable therapeutic potential is attributed not only to their differentiation and regenerative capacity, but also to the paracrine effect, underlying their immunomodulatory properties. MSCs’ secretome (i.e., cytokines, growth factors, and extracellular vesicles) is therefore increasingly discussed in the context of their ability to modulate inflammatory response and promote regeneration. There is evidence that 2D or 3D culturing conditions have an impact on the cells’ secretome, and here we aimed to compare the secretion of cytokines and growth factors in human MSCs from different sources cultured in 2D and 3D conditions and assess their effect on human macrophages polarization in vitro. Methods. MSCs were derived from human adipose tissue, bone marrow, gingiva, placenta, and umbilical cord, cultured as monolayers or as cell spheroids. Their cytokine profiles were analyzed, and data standardization was carried out using a z-score. Human peripheral blood mononuclear cells-derived macrophages were then treated with umbilical cord-derived MSCs’ conditioned media and their effect on macrophages polarization was assessed. Results. Our findings suggest that umbilical cord-derived MSCs’ conditioned media demonstrated the highest cytokine and growth factor levels and despite mostly pro-inflammatory cytokine profile were able to promote human macrophages’ polarization to anti-inflammatory M2 state and inhibit their polarization to pro-inflammatory M1 state. Conclusions. Umbilical cord-derived MSCs’ conditioned media hold great potential for therapeutic use, demonstrating significant anti-inflammatory effect on human macrophages.

Mesenchymal stromal cell derived extracellular vesicles as a therapeutic tool: immune regulation, MSC priming, and applications to SLE

“…While the secretome of MSC-EVs has shown to have high levels of IL-6 and IL-8 (27), a recent study showed that the conditioned media derived from umbilical cord (UC)-MSC promoted anti-inflammatory macrophages polarization. This despite a mostly proinflammatory profile of cytokines, though the authors have only investigated surface markers of M2 macrophages (28). MSC-EVs contain nucleic acids, which may bind to TLR7 and 9 as foreign nucleic acids and trigger a pro-inflammatory cascade.…”

Synergistic Hypoxia and Apoptosis Conditioning Unleashes Superior Mesenchymal Stem Cells Efficacy in Acute Graft-versus-Host-Disease

“…The polarization of macrophages from M1 to M2 phenotype was assessed by staining the cells with fluorochrome-conjugated anti-CD206 (BD Biosciences, USA) followed by fixation and permeabilization of the cells using an Intra Prep Permeabilization kit (Beckmann Coulter, USA) for intracellular staining. Further, the cells were stained with iNOS (Thermo Fisher Scientific, USA), CD68, and Arginase-1 (BD Biosciences, USA) for 30 minutes at room temperature and the cells were acquired using a DxFlex flow cytometer (Beckmann Coulter) to enumerate the M1 and M2 macrophages (31).…”