Corbu Arianna, Scaramozza Annarita, Badiali-DeGiorgi Lucilla, Tarantino Lucia, Papa Valentina, Rinaldi Rita, D'Alessandro Roberto, Zavatta Marcello, Laus Massimo, Lattanzi Giovanna, Cenacchi Giovanna
Clinical Department of Radiological and Histopathological Sciences, University of Bologna, Bologna, Italy.
Neurol Res. 2010 Feb;32(1):63-72. doi: 10.1179/174313209X385725.
Satellite cells (SCs) are skeletal muscle progenitor cells located between the basal lamina and the sarcolemma of muscle fibers. They are responsible for muscle growth and repair. In humans, aging results in the depletion of the SC population and in its proliferative activity, but not in its function. It has not yet been determined whether under conditions of massive muscle fiber death in vivo, the regenerative potential of SCs is totally or partially compromised in old muscle. No studies have yet tested whether advanced age is a factor that restrains the response of SCs to muscle denervation in humans; this is also due to difficulties in the isolation and in the culture of SCs from a small human surgery fragment. The aim of this study was to study in depth muscle regeneration analysing the SC ability of SCs to proliferate and differentiate in aging human patients.
In order to study in more detail the molecular mechanism, the proliferative and differentiative ability of aging SCs, we isolated SCs from aging human muscle biopsies and analysed their morphology by transmission electron microscopy and immunocytochemical analysis (antibodies against desmin, N-CAM and M-cadherin) and their capacity to grow and to expand in vitro. Moreover, in order to evaluate gene expression of myogenic regulatory factors Myf5, MyoD and myogenin (Myf4), RT-PCR was performed.
SCs isolated from aging human muscle biopsies and plated into favorable proliferation and differentiation conditions were able to proceed through the myogenic program and actively form myotubes, although taking longer than the young control sample. The RT-PCR analysis together with the ultrastructural SC features showed that the myogenic potential seemed to be compromised during the aging human muscle proliferation in vitro.
卫星细胞(SCs)是位于肌纤维基膜和肌膜之间的骨骼肌祖细胞。它们负责肌肉的生长和修复。在人类中,衰老导致卫星细胞数量减少及其增殖活性降低,但功能并未受损。目前尚未确定在体内大量肌纤维死亡的情况下,老年肌肉中卫星细胞的再生潜力是完全还是部分受损。尚无研究测试高龄是否是限制人类卫星细胞对肌肉去神经支配反应的因素;这也是由于从小的人体手术片段中分离和培养卫星细胞存在困难。本研究的目的是通过分析老年人类患者卫星细胞的增殖和分化能力,深入研究肌肉再生情况。
为了更详细地研究衰老卫星细胞的分子机制、增殖和分化能力,我们从老年人类肌肉活检组织中分离出卫星细胞,通过透射电子显微镜和免疫细胞化学分析(抗结蛋白、N-CAM和M-钙黏蛋白抗体)分析其形态,并分析它们在体外生长和扩增的能力。此外,为了评估生肌调节因子Myf5、MyoD和肌细胞生成素(Myf4)的基因表达,进行了逆转录聚合酶链反应(RT-PCR)。
从老年人类肌肉活检组织中分离出的卫星细胞,接种到有利于增殖和分化的条件下,能够进行生肌程序并积极形成肌管,尽管比年轻对照样本花费的时间更长。RT-PCR分析以及卫星细胞的超微结构特征表明,在老年人类肌肉体外增殖过程中生肌潜力似乎受到损害。