6533b862fe1ef96bd12c6a99

RESEARCH PRODUCT

G.P.199

subject

description

Our group has recently derived skeletal muscle from dermis-derived cells, by using an extracellular matrix that recreates the myogenic niche. After one week of differentiation, we observed isolated, twitching myotubes followed by spontaneous contractions of the entire tissue-engineered muscle construct. In vitro engineered myofibers expressed canonical markers, ultrastructure and electrophysiological characteristics of skeletal muscle. Interestingly, after one-month engineered muscle constructs showed progressive degradation of the myofibers concomitant with fatty infiltration, paralleling the natural course of muscular degeneration. However, we do not yet know how dermis-resident precursors are related with our isolated-myogenic precursors, a critical point for extrapolating our results to the human system. Knowing that dermal cells and muscle cells share a common embryonic origin from somite-derived dermomyotome and taking into account that there are different types of cells within the skin that have myogenic potential, our main objective is to identify and characterize the origin of murine dermal subpopulation with a myogenic potential, hypothesizing that may be (i) satellite cells from the Panniculus carnosus, (ii) dermomyotome-derived adult stem cells, (iii) perivascular cells and/or (iv) neural crest-derived precursor cells. For this end, lineage tracing experiments [(i) Pax3-GFP, (ii) Pax7CE, (iii) Myf5-Cre, (iv) CSPG4-Cre, and (v) Sox10-Cre] combined with FACS strategies and cellular differentiation assays have been developed. Our results show a high contribution of Myf5+ cells, a low contribution of Cspg4+ cells, and no contribution of Sox10+ cells to dermis-derived myogenic precursor cell subset. In principle, that means that dermomyotome-derived but not neural crest-derived cells are involved in subsequent engineered muscle differentiation.