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

来自猪和牛的骨骼肌细胞的体外成肌作用，目的是将细胞应用于肉类生产641020111297。利用猪和牛骨骼肌中的肌原细胞，对家畜进行了体外生肌试验。1)采用酶解聚和差异性细胞黏附相结合的方法制备猪和牛成肌细胞(SMC和BMC)，并用细胞松弛素B纯化。2)细胞内CK-MM和肌球蛋白鉴定的成肌细胞(Mb)的免疫细胞化学强度并确定其发育阶段。3)SMC和BMC在含有10%FCS和1.5%SFE的D-M培养液中生长显著。SMC可通过连续传代维持，形成肌管的能力较弱。4)体外建立了克隆的SMC，适合于研究成肌细胞的分化。5)需要胎牛血清来促进Mb的增殖，并将未成熟的Mb分化为Mb。从假定的Mb转变为不成熟的Mb，SFE是不可或缺的。6)在含有10%FCS和1.5%SFE的D-M培养液中加入Ca~(++)可使Mt和肌纤维(肌细胞：MC)形成。7)Mt和Mc的大规模培养因细胞载体材料的缺陷和融合促进因子的缺陷而不能成功。这些结果表明，家畜体外肌肉发生的进一步研究有可能与未来的肉类生产相联系。

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.