3D Culture and Interferon-γ Priming Modulates Characteristics of Mesenchymal Stromal/Stem Cells by Modifying the Expression of Both Intracellular and Exosomal microRNAs

Abstract

Mesenchymal stromal/stem cells (MSCs) have emerged as a therapeutic tool in regenerative medicine. Recent studies have shown that exosome (EXO)-derived microRNAs (miRNAs) play a crucial role in mediating MSC functions. Additionally, intracellular miRNAs have been found to regulate MSC therapeutic capacities. However, the molecular mechanisms underlying miRNA-mediated MSC effects are not fully understood. We used 3D culture and IFN-γ to prime/enhance the MSC therapeutic effects in terms of functional miRNAs. After priming, our analysis revealed stable variations in intracellular miRNA among the MSC biological replicates. Conversely, a significant variability of miRNA was observed among EXOs released from biological replicates of the priming treatment. For each priming, we observed distinct miRNA expression profiles between the MSCs and their EXOs. Moreover, in both types of priming, gene ontology (GO) analysis of deregulated miRNAs highlighted their involvement in tissue repair/regeneration pathways. In particular, the 3D culture enhanced angiogenic properties in both MSCs and EXOs, while IFN-γ treatment enriched miRNAs associated with immunomodulatory pathways. These findings suggest that 3D culture and IFN-γ treatment are promising strategies for enhancing the therapeutic potential of MSCs by modulating miRNA expression. Additionally, the identified miRNAs may contribute to understanding the molecular mechanisms underlying the miRNA-mediated therapeutic effects of MSCs.

Thermostable Basic Fibroblast Growth Factor Enhances the Production and Activity of Human Wharton’s Jelly Mesenchymal Stem Cell-Derived Extracellular Vesicles

“…WJ-MSCs possess various differentiation, self-renewal, and paracrine regulatory functions [38,39]. Paracrine signaling and therapeutic efficacy may be affected because enhanced exosome release has also been reported in WJ-MSC-derived exosomes carrying bioactive molecules that influence various biological processes, including immunomodulation, tissue regeneration, and angiogenesis [11,40]. This study elucidated the effects of bFGF treatment on the production and characteristics of EVs derived from WJ-MSCs.…”

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

“…Bulati et al were able to differentially cluster between IFN-g primed and 3D spheroid cultured MSCs by looking at their miRNA EVs (179). Many studies have shown that MSCs have increased immunomodulatory potential after 3D spheroid culture (159)(160)(161).…”

Extracellular Vesicles in Lung Cancer: Implementation in Diagnosis and Therapeutic Perspectives

“…Most preclinical studies involving MSC-EV therapy are based on vesicles produced by MSCs [ 3 , 64 , 65 ]. Moreover, to potentiate the functional activity of MSC-EVs, the strategy of priming/preconditioning their cells of origin was explored by using chemicals, cytokines, and growth factors, as well as specific culture conditions [ 3 , 64 , 66 , 67 , 68 , 69 ]. For instance, human MSC-EVs produced after stimulation with dimethyloxaloylglycine further stimulated angiogenesis through the Akt/mTOR pathway to enhance bone healing [ 70 ].…”

“…Furthermore, several studies explored the effects of inflammatory priming on MSC-EVs, revealing distinct EV functions compared to other priming conditions. For instance, it was recently demonstrated that EVs derived from IFN-γ-primed MSCs have improved immunomodulatory properties compared to the 3D culture priming of MSC-EVs, which instead showed enhanced angiogenic properties [ 66 ]. In this scenario, the yield, size, and surface marker composition of MSC-derived EVs exhibited substantial variations with various priming treatments, and it is intriguing to understand how the EV content and their beneficial properties can be modulated.…”