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

从培养的骨骼卫星细胞中分离的肌肉蛋白的利用的基础研究

基本信息

批准号：
63560275
负责人：
IKEUCHI Yoshihide
金额：
$ 1.02万
依托单位：
Niigata University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63560275/
关键词：
Satellite cell Growth factor Skeletal muscle Muscle fiber Regeneration 増殖因子筋肉蛋白質再生

项目摘要

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.

进行了一项研究，以实现油牛成纤维细胞生长因子对骨骼肌卫星细胞增殖的影响1。我们已经检查了使用涉及硫酸铵分馏的方法从牛脑组织中纯化脑成纤维细胞生长因子（脑FGF），然后在CM-Sephadex C-50上进行色谱法，Sephadex G-75和第二CM-Sephadex C-50。从第二CM-Sephadex C-50柱在0.35-0.4m NaCl中洗脱的分数在刺激BALB/C3T3细胞中的DNA合成方面有效。当通过SDS聚丙烯 - 酰胺凝胶电泳分析时，它主要由12,000个分子量的半肽组成。合并的数据（分子量和生物学活性）表明，使用此过程分离的分数包含大量由Gostodarowicz等人纯化的脑FGF（1978）.2。从成熟的大鼠培养骨骼肌卫星细胞，并用分离的脑FGF在体外进行治疗。形态学观察结果和定量数据表明大脑FGF增加了核的数量。新的核似乎是由卫星细胞增殖和其中一些融合而产生的。但是，新核的一部分源自在培养物中污染的成纤维细胞的生长。在15 0-250 ng/ml的范围内，当诱导卫星细胞分化时，该因子促进了增殖水平和随后的肌管形成水平。但是，卫星细胞的生物学活性比我们想象的要低。这可能是由于FGF制备中存在的杂质。