Exosomes secreted by human-induced pluripotent stem cell-derived mesenchymal stem cells attenuate limb ischemia by promoting angiogenesis in mice

Abstract

Introduction‘Patient-specific’ induced pluripotent stem cells (iPSCs) are attractive because they can generate abundant cells without the risk of immune rejection for cell therapy. Studies have shown that iPSC-derived mesenchymal stem cells (iMSCs) possess powerful proliferation, differentiation, and therapeutic effects. Recently, most studies indicate that stem cells exert their therapeutic effect mainly through a paracrine mechanism other than transdifferentiation, and exosomes have emerged as an important paracrine factor for stem cells to reprogram injured cells. The objective of this study was to evaluate whether exosomes derived from iMSCs (iMSCs-Exo) possess the ability to attenuate limb ischemia and promote angiogenesis after transplantation into limbs of mice with femoral artery excision.MethodsHuman iPSCs (iPS-S-01, C1P33, and PCKDSF001C1) were used to differentiate into iMSCs in a modified one-step method. iMSCs were characterized by flow cytometry and multipotent differentiation potential analysis. Ultrafiltration combined with a purification method was used to isolate iMSCs-Exo, and transmission electron microscopy and Western blotting were used to identify iMSCs-Exo. After establishment of mouse hind-limb ischemia with excision of femoral artery and iMSCs-Exo injection, blood perfusion was monitored at days 0, 7, 14, and 21; microvessel density in ischemic muscle was also analyzed. In vitro migration, proliferation, and tube formation experiments were used to analyze the ability of pro-angiogenesis in iMSCs-Exo, and quantitative reverse-transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay were used to identify expression levels of angiogenesis-related molecules in human umbilical vein endothelial cells (HUVECs) after being cultured with iMSCs-Exo.ResultsiPSCs were efficiently induced into iMSC- with MSC-positive and -negative surface antigens and osteogenesis, adipogenesis, and chondrogenesis differentiation potential. iMSCs-Exo with a diameter of 57 ± 11 nm and expressed CD63, CD81, and CD9. Intramuscular injection of iMSCs-Exo markedly enhanced microvessel density and blood perfusion in mouse ischemic limbs, consistent with an attenuation of ischemic injury. In addition, iMSCs-Exo could activate angiogenesis-related molecule expression and promote HUVEC migration, proliferation, and tube formation.ConclusionImplanted iMSCs-Exo was able to protect limbs from ischemic injury via the promotion of angiogenesis, which indicated that iMSCs-Exo may be a novel therapeutic approach in the treatment of ischemic diseases.Electronic supplementary materialThe online version of this article (doi:10.1186/scrt546) contains supplementary material, which is available to authorized users.

Beneficial effects of exosomes secreted by cardiac-derived progenitor cells and other cell types in myocardial ischemia

“…Besides iPSC exosomes, exosomes secreted by iPSC-derived MSCs have been investigated. The latter improved local angiogenesis in a limb ischemia mouse model and attenuated hepatic IR injury possibly via anti-inflammatory, anti-oxidant, and anti-apoptotic activities (74,75). These findings exemplify the therapeutic potential of iPSC exosomes and EVs from iPSC-derived MSCs.…”

Extracellular Vesicles Secreted by Human Urine-Derived Stem Cells Promote Ischemia Repair in a Mouse Model of Hind-Limb Ischemia

“…The mice were then randomly assigned to 4 different groups (n=15 for each group), which were intramuscularly injected with 100 μL of PBS (the PBS group), 1×10 6 USCs in 100 μL of PBS (the USC group), or USCs-EVs in 100 μL of PBS (the USCs-EVs group) via four sites of the gracilis muscle in the medial thigh as previously descried [25,26]. EVs secreted by induced pluripotent stem cell-derived MSCs (iMSCs-EVs) ameliorated limb ischemia in our previous study [27]; hence, mice injected with iMSCs-EVs in 100 μL of PBS (the iMSCs-EVs group) were used as a positive control. The concentration used in both the USCs-EVs and iMSCs-EVs groups was 2×10 10 particles/mL.…”

Mifepristone Inhibits the Migration of Cervical Cancer Cells by Inhibiting Exocrine Secretion

“…The scratch wound assay was used to analyze the migration ability of HeLa cells as described [17]. In brief, 6 × 10 3 cells were seeded into 24-well plates and cultured at 37 ° C for 18 h. Then the confluent monolayer cells were scratched by a 200 µL pipet tip.…”