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Myogenesis in vitro by skeletal muscle cells from swine and bovine for the purpose of applying the cells to meat production

猪和牛骨骼肌细胞的体外肌生成，旨在将细胞应用于肉类生产

基本信息

批准号：
02806049
负责人：
YAMAGUCHI Takahiro
金额：
$ 1.34万
依托单位：
Tohoku 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：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

Myogenesis in vitro by skeletal muscle cells from swine and bovine for the purpose of applying the cells to meat production 641020111297. Takahiro Yamaguchi, Tohoku University, Faculty of Agriculture, Assistant.Myogenests in vitro of domestic animals was examined by using myogenic cells from skeletal muscles of swine and bovine. Several findings were defined by the serial studies as follows :1)Swine and bovine myogenic cells(sMc and bMc)were prepared by the combination method of enzyme disaggregation and differential cell adhesion, and purified by cytochalasin B.2)Immunocytochemical intensity of intracellular CK-MM and myosin identified myoblasts(Mb)and defined their developmental stages.3)The growth of sMc and bMc was prominent in D-M medium with 10% FCS and 1.5% SFE. sMc could maintain by serial passage and had less ability to form myotubes(Mt). bMc formed many Mt although the cells could not maintain by passages.4)Cloned sMc were established in vitro and were suitable for studying the differentiation of myoblasts.5)FCS was needed to progress the proliferation of presumptive Mb and differentiate immature Mb into Mb. SFE was indispensable to transition from presumptive Mb into immature Mb. Active substances of SFE was contained in the fraction of 30-100 kd.6)The formation of Mt and muscle fibers(muscle cells : Mc)with bMc was accomplished by an addition of ca++ to D-M medium supplemented with 10% FCS and 1.5% SFE.7)The mass culture of Mt and Mc could not succeed because of the defective cell-carrier materials and the impaired fusion-promoting factors.The methods for Mb culture and Mt formation of swine and bovine established in this study can apply to medical and developmental biological research of myogenesis. The results point out the possibilities that further investigation on myogenesis in vitro domestic animals is connected to the meat production In future.

猪和牛骨骼肌细胞体外成肌作用及其在肉制品生产中的应用641020111297。利用猪和牛的骨骼肌的肌原性细胞，研究了家畜的体外肌原性。本研究的主要结果如下：1）采用酶解结合差速贴壁的方法制备了猪和牛的成肌细胞（sMc和bMc），并经细胞松弛素B纯化。2）细胞内CK-MM和肌球蛋白的免疫细胞化学强度鉴定了成肌细胞（Mb），并确定了它们的发育阶段。sMc可连续传代，但形成肌管的能力较弱。4）在体外建立了克隆的sMc，适合于研究成肌细胞的分化; 5）需要FCS来促进推定Mb的增殖和未成熟Mb向Mb的分化。SFE是从推定Mb向未成熟Mb过渡的必要条件。SFE的活性物质主要存在于30-100 kd的组分中。6）肌纤维和肌浆的形成（肌肉细胞：在含10%FCS和1.5%SFE的D-M培养基中加入Ca ~（++），使Mt和Mc与bMc融合。本研究建立的猪和牛肌红蛋白培养和肌红蛋白形成的方法可用于肌发生的医学和发育生物学研究。这些结果为进一步研究家畜离体肌肉发生与肉品生产的关系提供了可能。