Dimethyloxalylglycine, a small molecule, synergistically increases the homing and angiogenic properties of human mesenchymal stromal cells when cultured as 3D spheroids

Abstract

Strategies aiming at increasing the survival and paracrine activity of human mesenchymal stromal cells (MSCs) are of utmost importance to achieve the full therapeutic potential of these cells. Herein, we propose both physical and biochemical strategies to enhance the survival, homing, angiogenic, and immunomodulatory properties of MSCs in vitro. To that purpose, we compared the effect of exposing either 2D monolayer or 3D spheroids of MSCs to (i) hypoxia (2% O 2 ) or to (ii) a hypoxicmimetic small molecule, dimethyloxalylglycine (DMOG), with cells cultured at 21% O 2 . 3D-cultured MSC spheroids evidenced higher survival upon exposure to oxidative stress and expressed higher levels of factors involved in tissue repair processes, namely tumor necrosis factor-stimulated gene-6, matrix metalloproteinase-2, and vascular endothelial growth factor. MSCs cultured as 3D spheroids and further exposed to hypoxia or hypoxic-mimetic conditions provided by DMOG synergistically favored the expression of the cell surface marker C-X-C chemokine receptor type-4, involved in homing processes to injured tissues, and adhesion to extracellular matrix components as fibronectin. These results highlight the role of ex vivo preconditioning approaches, presenting a novel strategy that combine biochemical stimuli with 3D spheroid organization of MSCs to maximize their tissue regeneration potential.

High throughput screening of mesenchymal stromal cell morphological response to inflammatory signals for bioreactor-based manufacturing of extracellular vesicles that modulate microglia

“…Myriad biochemical and biomaterial MSC priming approaches have been documented and were not explored here [ 22 ]. For instance, small molecules have shown promise in priming MSCs and are amenable to screening [ 107 ]. However, our morphological profiling approach remains a generalizable platform for screening the effects of changes in manufacturing conditions (e.g., media, substrate, storage) with various cell types (e.g., iPSCs, NSCs).…”

Functional Mechanism of Bone Marrow-Derived Mesenchymal Stem Cells in the Treatment of Animal Models with Alzheimer’s Disease: Inhibition of Neuroinflammation

“… 25 , 63 Chemokine receptor CXCR4 is involved in the homing processes to injured tissues. 64 The expression of CXCR4, CCR2, Nrf2 and HIF-1α is upregulated in the MSCs, which mediates the rescue of learning and memory function in Aβ-injected rats. 62 When anti-apoptotic microRNA Let-7f-5p is used in APP/PS1 transgenic mice, the prolonged survival of BMMSCs can increase the therapeutic effect of BMMSCs.…”

Pretreated Mesenchymal Stem Cells and Their Secretome: Enhanced Immunotherapeutic Strategies

“…Compared with 2D culture, spheroid MSC-derived exosomes indicated enhanced immunomodulatory effects by restoring the reactive Th17/Treg balance in inflamed periodontal tissues and experimental colitis [ 110 ]. MSC spheroids in combination with hypoxia or hypoxic-mimetic condition provided by dimethyloxalylglycine synergistically increased TSG-6, MMP-2 and VEGF involved in tissue repair processes [ 111 ].…”