Identification of Satellite Cells in Skeletal Muscle

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  • Oka Kazuyuki
    Department of Biology, Toho University School of Medicine
  • Sakamoto Miki
    Faculty of Rehabilitation, School of Allied Health Sciences, Kitasato University
  • Nakamura Masaki
    Department of Biology, Toho University School of Medicine
  • Muro Masuo
    Exercise Physiology Lab. Toho University School of Medicine

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<p>  Satellite cells are the stem cells of skeletal muscle and they play an important role in the regeneration process. Understanding the activities of satellite cells well is essential to investigate the regeneration including hypertrophy, but it is difficult to distinguish satellite cells from myonuclei using a light microscope because the cytoplasm is extremely small. Immunohistochemical markers to identify satellite cells are reported but no general consensus on this has been reached among researchers. A transmission electron microscope (TEM) is the only definitive identification method for satellite cells even today. However, the observation area of the TEM section is limited and it requires a tremendous effort to make the specimen for TEM. In order to overcome these difficulties of TEM, we applied a scanning electron microscope (SEM). The goal of this study is to verify the appropriateness based on the satellite cell identification methods by SEM. </p><p>  In this study, the soleus muscles of 3-week-old and 12-week-old female ICR mice were used. Paraffin section were made and after sputter coating with Pt-Pd, the specimens were observed with SEM. Randomly the nuclei located inside the basal lamina were observed and the number of satellite cell nuclei per total number of nuclei was calculated as the rate of satellite cells in each animal. The ratio of satellite cells was compared with the previously published data of TEM. </p><p>  The rate of satellite cells was 7.5% on average in 12-week-old soleus muscle and it was 11.7% in 3-week-old soleus muscle. That is, the rate was significantly higher in young animals (P<0.0001). This age-related decreasing tendency coincided with the results in rat soleus muscle using TEM (Gibson et al. 1983). The rate of satellite cells in our experiments was almost the same as the TEM results in mouse soleus muscle (Snow 1981). </p><p>  Making specimens for SEM is easy compared to the specimens for TEM and, therefore, SEM is certain to be used widely to identify satellite cells in the future.</p>

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