Human Heart Explant-Derived Extracellular Vesicles: Characterization and Effects on the In Vitro Recellularization of Decellularized Heart Valves

Abstract

Extracellular vesicles (EVs) are particles released from different cell types and represent key components of paracrine secretion. Accumulating evidence supports the beneficial effects of EVs for tissue regeneration. In this study, discarded human heart tissues were used to isolate human heart-derived extracellular vesicles (hH-EVs). We used nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM) to physically characterize hH-EVs and mass spectrometry (MS) to profile the protein content in these particles. The MS analysis identified a total of 1248 proteins. Gene ontology (GO) enrichment analysis in hH-EVs revealed the proteins involved in processes, such as the regulation of cell death and response to wounding. The potential of hH-EVs to induce proliferation, adhesion, angiogenesis and wound healing was investigated in vitro. Our findings demonstrate that hH-EVs have the potential to induce proliferation and angiogenesis in endothelial cells, improve wound healing and reduce mesenchymal stem-cell adhesion. Last, we showed that hH-EVs were able to significantly promote mesenchymal stem-cell recellularization of decellularized porcine heart valve leaflets. Altogether our data confirmed that hH-EVs modulate cellular processes, shedding light on the potential of these particles for tissue regeneration and for scaffold recellularization.

The secretome from embryonic stem cell cardiomyogenesis: Same signals, different cellular feedbacks

“…Previously, our group verified that many proteins secreted at the final stages of hESC cardiac differentiation (D8/D9 and D15) were related to the cardiovascular system development (Robert et al, 2020). However, here, we demonstrated that CMd8 and Several studies showed the beneficial effects of using cell secretome in in vivo and in vitro investigations, assaying different types of cells and disease models (Ellison-Hughes & Madeddu, 2017;Ferreira et al, 2018;Leitolis, Suss et al, 2019;Li et al, 2019;S. R. Park et al, 2018;Pinho, Cibrão, Silva, Monteiro, & Salgado, 2020;Reus et al, 2016).…”

“…Several studies showed the beneficial effects of using cell secretome in in vivo and in vitro investigations, assaying different types of cells and disease models (Ellison‐Hughes & Madeddu, 2017; Ferreira et al, 2018; Leitolis, Suss et al, 2019; Li et al, 2019; S. R. Park et al, 2018; Pinho, Cibrão, Silva, Monteiro, & Salgado, 2020; Reus et al, 2016). Since these CM have a complex and unique composition, which varies according to cell type, donor, age, and culture conditions, the mechanisms that lead to the responses observed by each of these different CM are not yet fully understood.…”

“…Park et al, 2016;Ranganath, Levy, Inamdar, & Karp, 2012;Sharma et al, 2017;Zhang et al, 2015). Previously, our group demonstrated the potential of the secretome obtained from cardiac resident stromal cells in stimulating the proliferation and differentiation of H9c2 cells (Reus, Robert, Da Costa, de Aguiar, & Stimamiglio, 2016) and that the extracellular vesicles derived from human adult cardiac tissue could induce endothelial cell proliferation and angiogenesis (Leitolis, Suss et al, 2019). Despite the progress in this field, research remains focused on discovering different cell sources and strategies for CM collection with the aiming of finding an approach that allows the secretion of a set of molecules able to stimulate cardiac cell proliferation and/or differentiation.…”

“…The secretome from different cell sources, such as MSCs and cardiac progenitor cells (CPCs), were able to stimulate, among others, the angiogenesis of endothelial cells and the survival and proliferation of cardiomyocytes in vitro, in addition to improving cardiac function and stimulating resident cardiac progenitors after myocardial infarction in vivo (Balbi et al, 2019; Li et al, 2019; C. Park et al, 2016; Ranganath, Levy, Inamdar, & Karp, 2012; Sharma et al, 2017; Zhang et al, 2015). Previously, our group demonstrated the potential of the secretome obtained from cardiac resident stromal cells in stimulating the proliferation and differentiation of H9c2 cells (Reus, Robert, Da Costa, de Aguiar, & Stimamiglio, 2016) and that the extracellular vesicles derived from human adult cardiac tissue could induce endothelial cell proliferation and angiogenesis (Leitolis, Suss et al, 2019). Despite the progress in this field, research remains focused on discovering different cell sources and strategies for CM collection with the aiming of finding an approach that allows the secretion of a set of molecules able to stimulate cardiac cell proliferation and/or differentiation.…”

Cardiomyogenesis Modeling Using Pluripotent Stem Cells: The Role of Microenvironmental Signaling

“…Through the analysis of the ECM composition, soluble factors and EVs derived from cell and tissue cultures, we can search for essential signals and understand regulatory networks underlying human heart development. Regarding this issue, genomic and proteomic studies have provided important contributions (Reus et al, 2016; Wolling et al, 2018; Leitolis et al, 2019; Pereira et al, 2019). In the next sections, we will describe approaches to study the composition of cardiac secretome and discuss strategies that use the extracellular signals to induce PSC cardiac differentiation and maturation of PSC-CM.…”