Exosomes of human placenta-derived mesenchymal stem cells stimulate angiogenesis

Abstract

BackgroundThe therapeutic potential of mesenchymal stem cells (MSCs) may be attributed partly to humoral factors such as growth factors, cytokines, and chemokines. Human term placental tissue-derived MSCs (PlaMSCs), or conditioned medium left over from cultures of these cells, have been reported to enhance angiogenesis. Recently, the exosome, which can transport a diverse suite of macromolecules, has gained attention as a novel intercellular communication tool. However, the potential role of the exosome in PlaMSC therapeutic action is not well understood. The purpose of this study was to evaluate PlaMSC-derived exosome angiogenesis promotion in vitro and in vivo.MethodsMSCs were isolated from human term placental tissue by enzymatic digestion. Conditioned medium was collected after 48-h incubation in serum-free medium (PlaMSC-CM). Angiogenic factors present in PlaMSC-CM were screened by a growth factor array. Exosomes were prepared by ultracentrifugation of PlaMSC-CM, and confirmed by transmission electron microscopy, dynamic light scattering, and western blot analyses. The proangiogenic activity of PlaMSC-derived exosomes (PlaMSC-exo) was assessed using an endothelial tube formation assay, a cell migration assay, and reverse transcription-PCR analysis. The in-vivo angiogenic activity of PlaMSC-exo was evaluated using a murine auricle ischemic injury model.ResultsPlaMSC-CM contained both angiogenic and angiostatic factors, which enhanced endothelial tube formation. PlaMSC-exo were incorporated into endothelial cells; these exosomes stimulated both endothelial tube formation and migration, and enhanced angiogenesis-related gene expression. Laser Doppler blood flow analysis showed that PlaMSC-exo infusion also enhanced angiogenesis in an in-vivo murine auricle ischemic injury model.ConclusionsPlaMSC-exo enhanced angiogenesis in vitro and in vivo, suggesting that exosomes play a role in the proangiogenic activity of PlaMSCs. PlaMSC-exo may be a novel therapeutic approach for treating ischemic diseases.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0660-9) contains supplementary material, which is available to authorized users.

Angiogenic Effects of Human Dental Pulp and Bone Marrow-Derived Mesenchymal Stromal Cells and their Extracellular Vesicles

“…Komaki et al and Nakamura et al showed an equal induction of endothelial tube formation by CM and EVs derived from MSCs of human placental tissue and bone marrow. The associated EV-depleted CM did not affect this angiogenic step [24,62]. Similar results were also observed for human adipose-derived MSCs [63] and rat BM-MSCs [64].…”

Message in a Bottle: Upgrading Cardiac Repair into Rejuvenation

“…Likewise, EVs from UC-MSCs overexpressing the tissue matrix metalloproteinase inhibitor 2 (TIMP2) or the cardioprotective stromal-derived factor 1 alpha (SFD1a) have been shown to limit detrimental ventricular remodelling via the pro-survival Akt/Sfrp2 pathway and to inhibit apoptosis and autophagy of myocardial cells while sustaining local angiogenesis in preclinical rodent models of MI [200,201]. MSCs derived from human term placenta, also referred to as amniotic mesenchymal stromal cells (AMSCs), are well-known for their (immuno)modulatory properties [202,203]; EVs released by human term placenta-MSCs have been recently demonstrated exertion of relevant therapeutic effects as supporting new vessel development in vitro and in vivo [204], with nitric oxide (NO)-releasing polymer stimulation as a functional trigger of exosomal enrichment of pro-angiogenic VEGF and miR-126 [205]. Notably, placenta-MSCs and their derived EVs have been shown to counteract skewing of myoblasts to fibrogenic phenotype and collagen IV expression in the cardiac tissue of preclinical models of Duchenne musclar dystrophy, via targeted delivery of miR-29c [206].…”

Human Mesenchymal Stem Cells Promote Ischemic Repairment and Angiogenesis of Diabetic Foot Through Exosome miRNA-21-5p

“…To observe interactions between MSCs-exo and HUVECs, laser scanning confocal microscopy was used here. For uorescence staining of exosomes, MSCs-exo (8×10 9 ), miRNA-21-5p-KD-exo (8×10 9 ), and miRNA-21-5p-OE-exo (8×10 9 ) were respectively incubated with green uorescent dye PKH67 (MINI67-1KT, SIGMA) for 5 min at room temperature, followed by twice PBS wash using ultracentrifugation, as described previously 10 . Then, human umbilical vein endothelial cells (HUVECs) were co-cultured with either PBS (control), MSCs-exo, miRNA-21-5p-KD-exo or miRNA-21-5p-OE-exo for 4 hours at 37°C with 5% CO2.…”

“…Recent studies have reported that MSCs secrete exosomes, [9] which are membranous vesicles with diameter of 30-150 nm and density of 1.10 ~ 1.18 g/ml. [10] Exosomes contain various molecules including proteins, mRNA, and microRNA (miRNA), and these molecules can regulate the gene network of target cells as novel intercellular communicators. [11][12][13] During the past a few years, exosomes and exosome miRNA of MSCs in the angiogenesis had been increasingly attracted researcher's interests.…”