Abstract 15466: Bioactive Contents and Cardiovascular Regenerative Potential of Extracellular Vesicles Derived From Induced Pluripotent Stem Cells
Introduction: Extracellular vesicles (EVs) are tiny membrane-enclosed vesicles released by cells by membrane budding or exocytosis. The molecular profile of contents and regenerative potential of EVs derived from murine induced pluripotent stem cells (miPSCs) have not been investigated.
Hypothesis: We hypothesized that miPSC-derived EVs carry bioactive substances, including mRNA and miRNA, which would influence the biology and fate of target cells.
Methods: Murine iPSCs were generated using established protocols and EVs were isolated from culture supernatants by sequential centrifugation. Atomic force microscopy (AFM), flow cytometry, and real-time RT-PCR were used to characterize EV contents. Endothelial cells were exposed to miPSC-EVs in culture.
Results: AFM and dynamic light scattering showed that miPSC-EVs are homogenous spherical vesicles smaller than 100 nm (Fig, A). High-sensitivity flow cytometry confirmed the presence of several iPSC- specific markers along with typical exosomal markers (CD9, CD63 and CD81) on miPSC-EVs. miPSC-EVs were enriched in mRNAs, miRNAs and proteins from donor iPSCs as shown by real-time RT-PCR and mass spectroscopy, respectively. Moreover, miPSC-EVs contained transcripts regulating pluripotency, self-renewal and differentiation, including Oct-4, Nanog, GATA-4 as well as miR290-295 cluster (Fig, B,C). Importantly, several miRNAs were found to be higher in EVs when compared with parental miPSCs (Fig, B,C). Endothelial cells treated with miPSC-EVs exhibited greater proliferative, metabolic and angiogenic activities, and were more resistant to apoptosis.
Conclusions: Our data show, for the first time, that miPSC-derived EVs are natural nanocarriers capable of transferring bioactive contents to mature cells affecting target cell function as well as regenerative potential. We conclude that iPSC-EVs may represent safe therapeutic alternatives to whole cell-based therapy for cardiovascular repair.
Author Disclosures: M. Adamiak: None. S. Bobis-Wozowicz: None. E. Karnas: None. S. Kedracka-Krok: None. M. Sarna: None. G. Cheng: None. A. Samanta: None. M. Girgis: None. Z. Madeja: None. B. Dawn: None. E. Zuba-Surma: None.
- © 2015 by American Heart Association, Inc.