Radio Electric Conveyed Fields Directly Reprogram Human Dermal Skin Fibroblasts Toward Cardiac, Neuronal, and Skeletal Muscle-Like Lineages

Margherita Maioli, Salvatore Rinaldi, Sara Santaniello, Alessandro Castagna,
Gianfranco Pigliaru, Sara Gualini, Claudia Cavallini, Vania Fontani, and Carlo Ventura
Department of Biomedical Sciences, University of Sassari, Sassari, Italy
Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Bologna, Italy
Rinaldi Fontani Institute, Florence, Italy
§Cardiovascular Department, S. Orsola-Malpighi Hospital, University of Bologna, Bologna, Italy
Somatic cells can be directly reprogrammed to alternative differentiated fates without first becoming stem/
progenitor cells. Nevertheless, the initial need for viral-mediated gene delivery renders this strategy unsafe in
humans. Here, we provide evidence that exposure of human skin fibroblasts to a Radio Electric Asymmetric
Conveyer (REAC), an innovative device delivering radio electric conveyed fields at a radiofrequency of 2.4 GHz,
afforded remarkable commitment toward cardiac, neuronal, and skeletal muscle lineages. REAC induced the
transcription of tissue-restricted genes, including Mef2c, Tbx5, GATA4, Nkx2.5, and prodynorphin for cardiac
reprogramming, as well as myoD, and neurogenin 1 for skeletal myogenesis and neurogenesis, respectively.
Conversely, REAC treatment elicited a biphasic effect on a number of stemness-related genes, leading to early
transcriptional increase of Oct4, Sox2, cMyc, Nanog, and Klf4 within 6–20 h, followed by a downregulation at
later times. The REAC action bypassed a persistent reprogramming toward an induced pluripotent stem celllike
state and involved the transcriptional induction of the NADPH oxidase subunit Nox4. Our results show for
the first time the feasibility of using a physical stimulus to afford the expression of pluripotentiality in human
adult somatic cells up to the attainment of three major target lineages for regenerative medicine.