Extracellular vesicles from pluripotent stem cell-derived mesenchymal stem cells acquire a stromal modulatory proteomic pattern during differentiation

Abstract

Mesenchymal stem/stromal cells (MSCs) obtained from pluripotent stem cells (PSCs) constitute an interesting alternative to classical MSCs in regenerative medicine. Among their many mechanisms of action, MSC extracellular vesicles (EVs) are a potential suitable substitute for MSCs in future cell-free-based therapeutic approaches. Unlike cells, EVs do not elicit acute immune rejection, and they can be produced in large quantities and stored until ready to use. Although the therapeutic potential of MSC EVs has already been proven, a thorough characterization of MSC EVs is lacking. In this work, we used a label-free liquid chromatography tandem mass spectrometry proteomic approach to identify the most abundant proteins in EVs that are secreted from MSCs derived from PSCs (PD-MSCs) and from their parental induced PSCs (iPSCs). Next, we compared both datasets and found that while iPSC EVs enclose proteins that modulate RNA and microRNA stability and protein sorting, PD-MSC EVs are rich in proteins that organize extracellular matrix, regulate locomotion, and influence cell–substrate adhesion. Moreover, compared to their respective cells, iPSCs and iPSC EVs share a greater proportion of proteins, while the PD-MSC proteome appears to be more specific. Correlation and principal component analysis consistently aggregate iPSCs and iPSC EVs but segregate PD-MSC and their EVs. Altogether, these findings suggest that during differentiation, compared with their parental iPSC EVs, PD-MSC EVs acquire a more specific set of proteins; arguably, this difference might confer their therapeutic properties.

Highly Purified Human Extracellular Vesicles Produced by Stem Cells Alleviate Aging Cellular Phenotypes of Senescent Human Cells

“…Although iPSC‐derived EVs offer advantages in being produced in greater numbers and in stimulating cell metabolism, EVs from MSCs or other cell types that carry distinct cargos may be better suited for other medicinal applications. Recently, a proteomics study illustrated significant differences between human EVs produced by undifferentiated iPSCs and ones produced by iPSC‐derived isogenic MSCs . The former contains more cellular components of active cellular metabolism (consistent with our findings), whereas the latter acquires more stromal cell‐associated modulatory activities .…”

“…Recently, a proteomics study illustrated significant differences between human EVs produced by undifferentiated iPSCs and ones produced by iPSC-derived isogenic MSCs [47]. The former contains more cellular components of active cellular metabolism (consistent with our findings), whereas the latter acquires more stromal cell-associated modulatory activities [47]. One can envision that iPSC-derived EVs may be more efficient in medicinal applications to overcome cellular senescence, whereas EVs produced by various types of MSCs may be better suited to treat stromal disorders of collective tissues.…”

Human Pluripotent Stem Cell-Derived Extracellular Vesicles: Characteristics and Applications

“…78 EVs from iMSCs and undifferentiated iPSC-EVs were compared and their cargos were found to differ substantially. 79 While iPSC-EVs enclose proteins that modulate RNA and miRNA stability and protein sorting, iMSC-EVs enriched proteins that organize ECM and influence cell-substrate adhesions.…”

Proteomic Analysis of Exosomes from Adipose-Derived Mesenchymal Stem Cells: A Novel Therapeutic Strategy for Tissue Injury

“…La Greca et al identified 560 proteins from pluripotent stem cell-derived extracellular vesicles through proteomic analysis. Extracellular vesicles shared 37.32% of the proteins with parental cells and enriched with molecules relevant with immunity, extracellular matrix, and cell adhesion [39]. Otero-Ortega et al studied the proteomic signature of rat adipose-derived MSC-EVs in an experimental animal model of subcortical stroke.…”