Résumé :
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Communication n° 36. Multipotent adipose-derived stem (hMADS) cells have been isolated from the white adipose tissue removed from surgical scraps of infants undergoing surgery. The establishment and characterization of these cells have been described elsewhere (Rodriguez et al, submitted). Briefly, hMADS cells were selected as fast-adherent cells after seeding onto tissue culture plates the stromalvascular fraction cells. hMADS cells were shown to undergo more than 200 population doublings and to differentiate into cells of the adipogenic, osteogenic, angiogenic and myogenic lineages. The present work focuses on myogenic differentiation. hMADS were grown until nearly confluence and switched for myogenic differentiation medium. Labelling of KI67 protein indicated that hMADS cells exit cell cycle under these conditions. Many myogenic differentiation cell culture conditions were tested. hMADS cells aligned contiguously to each other, expressed myogenin but did not fuse into myotubes. hMADS cells were then grown in coculture with various myogenic cells. The cells we used were the murine C2C12 cells, primary mdx mouse myoblasts, primary human myoblasts from a control individual, and a myoblast cell line derived from a Duchene muscular dystrophy patient. Experimental culture conditions were optimized. Cocultures were grown in proliferative then differentiation media. hMADS cell nuclei were identified in myotubes and we present the expression analysis of early and late markers of interest for myogenic differentiation. Conclusions for hMADS cell myogenic differentiation capacities are discussed.
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