Interferon-γ and Hypoxia Priming Have Limited Effect on the miRNA Landscape of Human Mesenchymal Stromal Cells-Derived Extracellular Vesicles

• Published in: Frontiers in cell and developmental biology Vol. 8; p. 581436
• Main Authors: Peltzer, Juliette, Lund, Kyle, Goriot, Marie-Emmanuelle, Grosbot, Marion, Lataillade, Jean-Jacques, Mauduit, Philippe, Banzet, Sébastien
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers media 15.12.2020; Frontiers Media S.A
• Subjects: Biotechnology; Cell and Developmental Biology; Cellular Biology; exosome (vesicle); extracellular vesicles; Life Sciences; mesenchymal stem cells; mesenchymal stromal cells; miRNA; priming; exosome (vesicle); priming; miRNA; extracellular vesicles; mesenchymal stem cells; mesenchymal stromal cells
• ISSN: 2296-634X; 2296-634X
• DOI: 10.3389/fcell.2020.581436

Mesenchymal stromal cell (MSC)-based cell therapy has received great interest in regenerative medicine. Priming the cells during the culture phase can improve their efficacy and/or survival after injection. The literature suggests that MSC extracellular vesicles (EV) can recapitulate a substantial part of the beneficial effects of the cells they originate from, and that micro-RNAs (miRNAs) are important players in EV biological action. Here, our aim was to determine if two classical priming methods of MSC, interferon-gamma (IFNγ) and hypoxia (HYP), could modify their EV miRNA content. Human bone marrow MSCs (BM-MSCs) from five healthy donors were cultured with IFNγ or in HYP or in control (CONT) conditions. The conditioned media were collected after 48 h in serum-free condition and EV were isolated by ultracentrifugation. Total RNA was isolated, pools of CONT, IFN, and HYP cDNA were prepared, and a miRNA profiling was performed using RT-qPCR. Then, miRNAs were selected based on their detectability and measured on each individual EV sample. Priming had no effect on EV amount or size distribution. A set of 81 miRNAs was detected in at least one of the pools of EVs. They were measured on each individual sample; 41 miRNAs were detected in all samples. The principal component analysis (PCA) failed to discriminate the groups. HYP induced a significant decrease in EV hsa-miR-34a-3p content and IFN induced a significant increase in five miRNAs (hsa-miR-25-3p, hsa-miR-106a-5p, hsa-miR-126-3p, hsa-miR-451a, and hsa-miR-665). Taken together, we found only limited alterations in the miRNA landscape of MSC EV with a high inter-individual variability.