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在特定无血清体系中由成年大鼠卫星细胞形成功能性肌管

Functional myotube formation from adult rat satellite cells in a defined serum-free system.

作者信息

McAleer Christopher W, Rumsey John W, Stancescu Maria, Hickman James J

机构信息

NanoScience Technology Center, University of Central Florida, 12424 Research Parkway, Suite 400, Orlando, FL, 32826.

出版信息

Biotechnol Prog. 2015 Jul-Aug;31(4):997-1003. doi: 10.1002/btpr.2063. Epub 2015 Mar 4.

Abstract

This manuscript describes the development of a culture system whereby mature contracting myotubes were formed from adult rat derived satellite cells. Satellite cells, extracted from the Tibialis Anterior of adult rats, were grown in defined serum-free growth and differentiation media, on a nonbiological substrate, N-1[3-trimethoxysilyl propyl] diethylenetriamine. Myotubes were evaluated morphologically and immunocytochemically, using MyHC specific antibodies, as well as functionally using patch clamp electrophysiology to measure ion channel activity. Results indicated the establishment of the rapid expression of adult myosin isoforms that contrasts to their slow development in embryonic cultures. This culture system has applications in the understanding and treatment of age-related muscle myopathy, muscular dystrophy, and for skeletal muscle engineering by providing a more relevant phenotype for both in vitro and in vivo applications.

摘要

本手稿描述了一种培养系统的开发,通过该系统可从成年大鼠来源的卫星细胞形成成熟的收缩性肌管。从成年大鼠胫骨前肌中提取的卫星细胞,在限定的无血清生长和分化培养基中,在非生物底物N-1[3-三甲氧基甲硅烷基丙基]二乙三胺上生长。使用肌球蛋白重链(MyHC)特异性抗体对肌管进行形态学和免疫细胞化学评估,并使用膜片钳电生理学功能测量离子通道活性。结果表明,成年肌球蛋白同工型快速表达得以确立,这与它们在胚胎培养物中的缓慢发育形成对比。该培养系统通过为体外和体内应用提供更相关的表型,在理解和治疗与年龄相关的肌肉肌病、肌肉营养不良以及骨骼肌工程方面具有应用价值。

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