Immortalized pathological human myoblasts: towards a universal tool for the study of neuromuscular disorders
Immortalized pathological human myoblasts: towards a universal tool for the study of neuromuscular disorders
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DOI：
10.1186/2044-5040-1-34
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发表时间：
2011-01-01
期刊：
影响因子：
4.9
通讯作者：
Mouly, Vincent
中科院分区：
文献类型：
--
作者：
Mamchaoui, Kamel;Trollet, Capucine;Mouly, Vincent
背景资料：研究肌营养不良的病理生理学和治疗靶点一直受到人成肌细胞有限增殖能力的阻碍。从患者活检组织中分离可靠和稳定的永生化细胞系是研究病理机制（包括与肌肉老化相关的病理机制）和开发创新的基于基因、基于细胞或药理学的生物疗法的有力工具。使用端粒酶表达载体和细胞周期蛋白依赖性激酶4表达载体的转导，我们能够从患有各种神经肌肉疾病的患者中产生一系列永生化的人类肌肉干细胞系。来自Duchenne肌营养不良症、面肩肱肌营养不良症、眼咽肌营养不良症、先天性肌营养不良症和肢带型肌营养不良症2B型移植到免疫缺陷小鼠的再生肌肉中后，其增殖能力大大增加，并保持其体外和体内分化的潜力。需要营养不良细胞模型作为动物模型的补充，以评估细胞机制，如信号传导缺陷，或对治疗分子进行高通量筛选。这些研究已经在动物来源的细胞上进行了多年，并且将极大地受益于具有延长的增殖能力的人类细胞模型。此外，在体内评估这些细胞的再生能力的可能性扩展了它们的潜在用途。如本报告所述，来自几种不同神经肌肉疾病的创新细胞工具将允许调查这些疾病的病理生理学和评估新的治疗策略。
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.