TY - JOUR
T1 - Immortalized pathological human myoblasts
T2 - Towards a universal tool for the study of neuromuscular disorders
AU - Mamchaoui, Kamel
AU - Trollet, Capucine
AU - Bigot, Anne
AU - Negroni, Elisa
AU - Chaouch, Soraya
AU - Wolff, Annie
AU - Kandalla, Prashanth K.
AU - Marie, Solenne
AU - Di Santo, James
AU - St Guily, Jean L.
AU - Muntoni, Francesco
AU - Kim, Jihee
AU - Philippi, Susanne
AU - Spuler, Simone
AU - Levy, Nicolas
AU - Blumen, Sergiu C.
AU - Voit, Thomas
AU - Wright, Woodring E.
AU - Aamiri, Ahmed
AU - Butler-Browne, Gillian
AU - Mouly, Vincent
N1 - Funding Information:
We thank all the patients who provided the biopsies to establish the primary cultures. We also thank the MSG study group, particularly D. Furling for fruitful discussions and L. Dollé, S. Sandal, M. Oloko, S. Vasseur and M. Chapart for technical assistance. We thank Genosafe for help with the transductions and Steve Wilton for providing the antisense oligonucleotides. This work was supported by the MYORES Network of Excellence (contract 511978) and TREAT-NMD (contract LSHM-CT-2006-036825) from the European Commission 6th FP, MYOAGE (contract HEALTH-F2-2009-223576) from the Seventh FP, the ANR Genopath-INAFIB, the ANR MICRORNAS, MyoGrad (GK1631, German Research Foundation), the Duchenne Parent Project Netherlands, CNRS, INSERM, University Pierre and Marie Curie, AFM (Association Française contre les Myopathies) (including network grant #15123), the Jain Foundation, Parents Project of Monaco, and the European Parent Project.
PY - 2011/11/1
Y1 - 2011/11/1
N2 - Background: Investigations into both the pathophysiology and therapeutic targets in muscle dystrophies have been hampered by the limited proliferative capacity of human myoblasts. Isolation of reliable and stable immortalized cell lines from patient biopsies is a powerful tool for investigating pathological mechanisms, including those associated with muscle aging, and for developing innovative gene-based, cell-based or pharmacological biotherapies.Methods: Using transduction with both telomerase-expressing and cyclin-dependent kinase 4-expressing vectors, we were able to generate a battery of immortalized human muscle stem-cell lines from patients with various neuromuscular disorders.Results: The immortalized human cell lines from patients with Duchenne muscular dystrophy, facioscapulohumeral muscular dystrophy, oculopharyngeal muscular dystrophy, congenital muscular dystrophy, and limb-girdle muscular dystrophy type 2B had greatly increased proliferative capacity, and maintained their potential to differentiate both in vitro and in vivo after transplantation into regenerating muscle of immunodeficient mice.Conclusions: Dystrophic cellular models are required as a supplement to animal models to assess cellular mechanisms, such as signaling defects, or to perform high-throughput screening for therapeutic molecules. These investigations have been conducted for many years on cells derived from animals, and would greatly benefit from having human cell models with prolonged proliferative capacity. Furthermore, the possibility to assess in vivo the regenerative capacity of these cells extends their potential use. The innovative cellular tools derived from several different neuromuscular diseases as described in this report will allow investigation of the pathophysiology of these disorders and assessment of new therapeutic strategies.
AB - Background: Investigations into both the pathophysiology and therapeutic targets in muscle dystrophies have been hampered by the limited proliferative capacity of human myoblasts. Isolation of reliable and stable immortalized cell lines from patient biopsies is a powerful tool for investigating pathological mechanisms, including those associated with muscle aging, and for developing innovative gene-based, cell-based or pharmacological biotherapies.Methods: Using transduction with both telomerase-expressing and cyclin-dependent kinase 4-expressing vectors, we were able to generate a battery of immortalized human muscle stem-cell lines from patients with various neuromuscular disorders.Results: The immortalized human cell lines from patients with Duchenne muscular dystrophy, facioscapulohumeral muscular dystrophy, oculopharyngeal muscular dystrophy, congenital muscular dystrophy, and limb-girdle muscular dystrophy type 2B had greatly increased proliferative capacity, and maintained their potential to differentiate both in vitro and in vivo after transplantation into regenerating muscle of immunodeficient mice.Conclusions: Dystrophic cellular models are required as a supplement to animal models to assess cellular mechanisms, such as signaling defects, or to perform high-throughput screening for therapeutic molecules. These investigations have been conducted for many years on cells derived from animals, and would greatly benefit from having human cell models with prolonged proliferative capacity. Furthermore, the possibility to assess in vivo the regenerative capacity of these cells extends their potential use. The innovative cellular tools derived from several different neuromuscular diseases as described in this report will allow investigation of the pathophysiology of these disorders and assessment of new therapeutic strategies.
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U2 - 10.1186/2044-5040-1-34
DO - 10.1186/2044-5040-1-34
M3 - Article
C2 - 22040608
AN - SCOPUS:84862612146
SN - 2044-5040
VL - 1
JO - Skeletal Muscle
JF - Skeletal Muscle
IS - 1
M1 - 34
ER -