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1 Istituto Pasteur-Fondazione Cenci Bolognetti, Dipartimento di Fisiologia Umana e Farmacologia and Centro di Eccellenza BEMM, Universit La Sapienza Piazzale Aldo Moro 5, I-00185 Roma, Italy
2
Dipartimento di Istologia ed Embriologia medica and Centro di Eccellenza BEMM, Università ‘La Sapienza’, via A. Scarpa 14, 00161 Roma, Italy
3
Stem Cell Research Institute, Dibit, Istituto San Raffaele, Via Olgettina 58, I-20132 Milano, Italy
Mesoangioblasts are vessel-associated fetal stem cells that can be induced to differentiate into skeletal muscle, both in vitro and in vivo. Whether this is due to fusion or to transdifferentiation into bona fide satellite cells is still an open question, for mesoangioblasts as well as for other types of stem cells. The early steps of satellite cell myogenic differentiation involve MyoD activation, membrane hyperpolarization and the appearance of ACh sensitivity and gap junctional communication. If mesoangioblasts differentiate into satellite cells, these characteristics should be observed in stem cells prior to fusion into multinucleated myotubes. We have investigated the functional properties acquired by mononucleated green fluorescent protein (GFP)-positive mesoangioblasts co-cultured with differentiating C2C12 myogenic cells, using the patch-clamp technique. Mesoangioblasts whose membrane contacted myogenic cells developed a hyperpolarized membrane resting potential and ACh-evoked current responses. Dye and electrical coupling was observed among mesoangioblasts but not between mesoangioblasts and myotubes. Mouse MyoD was detected by RT-PCR both in single, mononucleated mesoangioblasts co-cultured with C2C12 myotubes and in the total mRNA from mouse mesoangioblasts co-cultured with human myotubes, but not in human myotubes or stem cells cultured in isolation. In conclusion, when co-cultured with muscle cells, mesoangioblasts acquire many of the functional characteristics of differentiating satellite cells in the absence of cell fusion, strongly indicating that these stem cells undergo transdifferentiation into satellite cells, when exposed to a myogenic environment.
(Received 25 June 2004;
accepted after revision 17 August 2004;
first published online 19 August 2004)
Corresponding author F. Grassi: Istituto Pasteur-Fondazione Cenci Bolognetti, Dipartimento di Fisiologia Umana e Farmacologia and Centro di Eccellenza BEMM, Università La Sapienza P.le A. Moro 5; I-00185 Roma, Italy. Email: francesca.grassi{at}uniroma1.it
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