Résumé :
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We recently generated immortalized human myoblast cell lines from control donors, by transduction of these cells with both telomerase (hTERT) and cyclin-dependent kinase 4 (CDK-4) expressing vectors. These cells maintain their potential to differentiate both in vitro and in vivo, and are not tumorigenic (Zhu, Mouly et al 2007). We have now generated human myoblast and fibroblast cell lines from various neuromuscular disorders using the same approach. Fibroblasts can be easily isolated and then converted by transduction of an inducible MyoD expression vector. Such converted fibroblasts have been used to assess exon skipping strategy for both DMD and dysferlin deficiency. We have also now generated 3 DMD myoblast cell lines with different mutations, which can be corrected by exon skipping for 2 of these (skipping exon 51 and 53 respectively), or by stop codon readthrough for the third one. We have also generated 5 different dysferlin myoblast cell lines with again a different set of mutations, and also an immortal cell line from a FSHD patient (2 D4Z4). Regarding OPMD, we have isolated immortal cell lines from 2 heterozygote patients, both from affected and non affected muscles, as well as a cell line from an homozygote patient. Finally, we have also generated a cell line from a patient suffering from a congenital muscular dystrophy due to a mutation in the nesprin gene. Their capacity to differentiate in vitro as well as their proliferative potential and telomere lengths were monitored for all these cell lines. Furthermore, their capacity to participate to in vivo regeneration was assessed for some of these cell lines, by injecting them into regenerating TA muscles of immunodeficient mice. All these cell lines represent useful tools to assess pathophysiology for diseases in which, although the mutation is known, the mechanisms leading to the pathology are still unclear, such as FSHD, OPMD, or nesprin deficiency. For diseases in which the pathological mechanism is at least partially known, such as DMD or dysferlin deficiency, they represent valuable tools to assess tailored or general therapeutic strategies, depending on the targeted mutation. More cell lines are currently being developed, and these cell lines can be shared with the scientific community on a collaborative basis.
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