Titre :
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Transplantation of purified myoblasts derived from embryonic stem cells (abstract : congrès international de Myologie, 2005)
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contenu dans :
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Auteurs :
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Congrès international de myologie 2005 (International Congress of Myology 2005; 9-13 mai 2005; Nantes, France) ;
Lapointe E ;
Ducharme ME ;
Camirand G ;
Kramer R ;
Tremblay J ;
Skuk D
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Type de document :
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Article
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Année de publication :
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2005
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Pages :
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p. 286
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Langues:
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Anglais
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Mots-clés :
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cellule souche embryonnaire
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colloque
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desmine
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différenciation cellulaire
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dystrophie musculaire de Duchenne
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dystrophine
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facteur de croissance
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IGF1
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METHODOLOGIE
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myoblaste
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prolifération cellulaire
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souris
;
thérapie cellulaire
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Résumé :
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Communication n° 199 Introduction : Duchenne muscular dystrophy (DMD) is characterized by the absence of the dystrophin muscle protein (427 kDa). Delivery of normal dystrophin gene by myoblast transplantation permits the long term restoration of this protein. One problem of this technique is the limited number of myoblasts that can be obtained from a non dystrophic donor. Objective : The differentiation of embryonic stem (ES) cells could provide an unlimited number of myogenic cells for grafts. Methods : Mouse ES cells were grown in a differentiation media with insulin-like growth factor-1 (IGF-1) and hepatocyte growth factor (HGF) to promote their differentiation into myoblasts. The percentage of ES cells differentiated into myogenic cells was determined by immunocytochemistry staining for desmin. After ES cell differentiation, myogenic cells were purified by magnetic affinity cell sorting (MACS) using an antibody against ?-7 integrin. Purified myoblasts were transplanted into the Tibialis anterior (TA) of mdx mice. Non-purified cells were also grafted as a control to evaluate the necessity of the purification procedure. Immunohistochemical stainings were performed to identify the dystrophin positive muscle fibres and an hematoxylin/eosin staining was done to detect the presence of tumours in transplanted muscles. Results : Addition of IGF-1 and HGF to the medium produced a 15% myoblast differentiation after three days. Without these growth factors, the ES cells remained partly undifferentiated after three days and were not positive for desmin. The expression of dystrophin in the grafted muscles showed that purified ES cells were able to proliferate and fuse in vivo. Purification of differentiated ES cells by MACS prevented the formation of tumours. Conclusion : This study showed that it is possible to obtain myoblasts from ES cells in vitro. Gene expression and further analysis will permit to improve the differentiation efficacy. This method, applied on human ES cells, would allow the treatment of DMD patients without the use of a specific donor for each patient.
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