Titre : | A proteomic investigation of human satellite cell senescence in vitro (abstract : congrès international de Myologie, 2005) |
contenu dans : | |
Auteurs : | Congrès international de myologie 2005 (International Congress of Myology 2005; 9-13 mai 2005; Nantes, France) ; Le Bihan M ; Bigot A ; Butler-Browne GS ; Mouly V ; Furling D ; Coulton G |
Type de document : | Article |
Année de publication : | 2005 |
Pages : | p. 93 |
Langues: | Anglais |
Mots-clés : | colloque ; culture cellulaire ; humain ; in vitro ; mitose ; muscle squelettique ; myoblaste ; prolifération cellulaire ; télomère ; vieillissement |
Résumé : |
Communication n° 571. Introduction : The proliferation of human satellite cells in vitro is limited by replicative senescence. For example, satellite cells isolated from newborns can achieve 55-60 divisions in vitro, while cells isolated from adults only make 15-25 divisions. As is the case for other human somatic cells, the proliferative capacity of satellite cells is limited by a mitotic clock determined by telomere length. However, the intricacies of the molecular networks that are associated with and control senescence remain to be described in detail. The identification of networks regulating cell senescence may prove to have direct relevance for example to understanding muscle ageing in vivo, the pathogenesis of myopathies and the efficacy of cell-based therapy for muscle disease. Objective : The aim of this study was to characterise differences in global patterns of protein expression observed during replicative senescence of human satellite cells in vitro. Methods : Human satellite cells harvested from a five-day old baby at autopsy were cultivated until they reached proliferative pre-senescence. Proliferating and fully differentiated cultures derived from cells at early passage and pre-senescence were analysed proteomically. Intracellular extracts and proteins secreted into culture supernatants were analysed. We utilised an integrated proteomic pipeline comprising Ciphergen protein arrays, Surface-Enhanced Laser Desorption and Ionisation (SELDI) mass spectrometry, 2D gel electrophoresis and Matrix Assisted Laser Desorption and Ionisation (MALDI) mass spectrometry. The strategy comprised several stages. 1) Protein extraction, 2) Biomarker Discovery, 3) Biomarker validation & 4) Biomarker Identification. Results : We discovered 21 intracellular biomarkers discriminating proliferating early passage from proliferating pre-senescent cells and 29 intracellular biomarkers discriminating differentiated early passage from differentiated pre-senescent cells. We will also present data for secreted proteins. Conclusions : The proteomic expression patterns of early passage and pre-senescent satellite cells are significantly different. The biomarkers identified will be investigated subsequently for their potential as regulators of cell senescence in vitro and also as markers of muscle ageing in vivo. |