Basic Study on the Utilization of Muscle Proteins Isolated from Cultured Skeletal Satellite Cells

本实验研究了牛成纤维细胞生长因子对骨骼肌卫星细胞增殖的影响.我们已经研究了从牛脑组织中纯化脑成纤维细胞生长因子（Brain FGF）的方法，该方法包括硫酸铵分级分离，然后在CM-Sephadex C-50、Sephadex G-75和第二个CM-Sephadex C-50上层析。在0.35-0.4M NaCl下从第二个CM-SephadexC-50柱洗脱的级分对Balb/c3 T3细胞中的DNA合成具有刺激活性。SDS聚丙烯酰胺凝胶电泳分析表明，其主要由分子量为12，000的多肽组成。结合的数据（分子量和生物活性）表明，使用该方法分离的级分含有大量的由Gospodarowicz等人纯化的脑FGF。（1978年）。从成熟大鼠中培养骨骼肌卫星细胞，并在体外用分离的脑FGF处理。形态学观察和定量数据表明，脑FGF增加核的数量。新的细胞核似乎是卫星细胞增殖和其中一些细胞融合的结果。但也有一部分新细胞核来源于培养物中污染的成纤维细胞的生长。在150 ~ 250 ng/ml范围内，该因子促进卫星细胞的增殖和肌管形成。然而，卫星细胞的生物活性比我们想象的要低得多。这可能是由于FGF制剂中存在杂质所致。

An investigation was undertaken to the effect oil bovine fibroblast growth factor on skeletal muscle satellite cell proliferation.1. We have examined to purify brain fibroblast growth factor (Brain FGF) from bovine brain tissue using this procedure involving ammonium sulfate fractionation, followed by chromatography on CM-Sephadex C-50, Sephadex G-75 and second CM-Sephadex C-50. The fraction eluting from second CM-Sephadex C-50 column at 0.35-0.4M NaCl was active in stimulating DNA synthesis in Balb/c3T3 cells. It mainly consisted of apolypeptide of 12,000 molecular weight when analyzed by SDS polyacryl-amide gel electrophoresis. The combined data ( molecular weight and biological activity ) suggested that the fraction isolated using this procedure contained a large quantity of brain FGF purified by Gospodarowicz et al.(1978).2. Skeletal muscle satellite cells were cultured from mature rats and were treated in vitro with the isolated brain FGF. Morphological observations and quantitative data indicated that the brain FGF increased the number of nuclei. The new nuclei seemed to result from satellite cell proliferation and from the fusion of some of them. But, a part of new nuclei originated from growth of fibroblast contaminated in cultures. In the range of 15 0-250 ng/ml , this factor promoted the level of proliferation and subsequent myotube formation when satellite cells were induced to differentiate. However, the biological activity for satellite cells was fairly lower than we had imagined. This may be due to impurities present in the FGF preparation.