**Extracellular Vesicles from Skeletal Muscle Cells E**ffi**ciently Promote Myogenesis in Induced Pluripotent Stem Cells**

**Denisa Baci 1,2, Maila Chirivì 1, Valentina Pace 1, Fabio Maiullari 3, Marika Milan 1, Andrea Rampin 1, Paolo Somma 4, Dario Presutti 1, Silvia Garavelli 5, Antonino Bruno 6, Stefano Cannata 7, Chiara Lanzuolo 8,9, Cesare Gargioli 7, Roberto Rizzi 8,9,\* and Claudia Bearzi 1,9,\***


Received: 19 April 2020; Accepted: 15 June 2020; Published: 23 June 2020

**Abstract:** The recent advances, offered by cell therapy in the regenerative medicine field, offer a revolutionary potential for the development of innovative cures to restore compromised physiological functions or organs. Adult myogenic precursors, such as myoblasts or satellite cells, possess a marked regenerative capacity, but the exploitation of this potential still encounters significant challenges in clinical application, due to low rate of proliferation in vitro, as well as a reduced self-renewal capacity. In this scenario, induced pluripotent stem cells (iPSCs) can offer not only an inexhaustible source of cells for regenerative therapeutic approaches, but also a valuable alternative for in vitro modeling of patient-specific diseases. In this study we established a reliable protocol to induce the myogenic differentiation of iPSCs, generated from pericytes and fibroblasts, exploiting skeletal muscle-derived extracellular vesicles (EVs), in combination with chemically defined factors. This genetic integration-free approach generates functional skeletal myotubes maintaining the engraftment ability in vivo. Our results demonstrate evidence that EVs can act as biological "shuttles" to deliver specific bioactive molecules for a successful transgene-free differentiation offering new opportunities for disease modeling and regenerative approaches.

**Keywords:** iPSC; extracellular vesicles; pericytes; skeletal muscle
