鲁西黑头羊肌肉细胞体外培养及诱导分化研究  

Culture in vitro and Induced Differentiation of Muscle Cells in Luxi Blackhead Sheep

作　　者：刘昭华[1] 王可[1] 崔绪奎[1] 靳青[1] 朱荣生[1] 王建英[1] 孟宪锋[1] 张冬梅 谭秀文[1] Liu Zhaohua;Wang Ke;Cui Xukui;Jin Qing;Zhu Rongsheng;Wang Jianying;Meng Xianfeng;Zhang Dongmei;Tan Xiuwen(Institute of Animal Science and Veterinary Medicine,Shandong Academy of Agricultural Sciences/Shandong Key Laboratory of Animal Disease Control and Breeding,Jinan 250100,China;Zoucheng Development Center of Animal Husbandry,Zoucheng 273500,China)

机构地区：[1]山东省农业科学院畜牧兽医研究所/山东省畜禽疫病防治与繁育重点实验室,山东济南250100 [2]山东省邹城市畜牧业发展中心,山东邹城273500

出　　处：《山东农业科学》2021年第10期116-122,共7页Shandong Agricultural Sciences

基　　金：山东省农业科学院农业科技创新工程项目(CXGC2021A14);国家现代农业产业技术体系(肉羊)项目(CARS-39);山东省现代农业产业技术体系项目(SDAIT-10-02)。

摘　　要：细胞体外培养是研究肌肉生长发育最有利的工具,迄今为止,肉羊肌肉细胞体外培养体系一直不成熟,有关细胞生长规律以及分化后超微结构的报道较少。本研究利用酶消化法培养鲁西黑头羊肌肉细胞,通过流式细胞仪对细胞进行肌源性鉴定,通过绘制生长曲线对细胞生长规律进行探索,利用分化培养液对细胞进行成肌诱导,并利用扫描电镜和透射电镜对其超微结构进行观察。结果发现,培养的鲁西黑头羊肌肉细胞纯度达到95%以上。肌肉细胞经历潜伏期、指数生长期、平台期和衰退期,其中平台期较长,肌肉细胞在这一时期自发分化。诱导分化后细胞表面伸出丝足,质膜分布有很多空隙孔;线粒体十分丰富,内嵴发达、致密;糙面内质网数量较多,表面核糖体清晰可见;胞浆内散在分布大量的脂滴和糖原;肌原纤维束致密,排列规则。综之,本研究成功构建了鲁西黑头羊肌肉细胞培养体系,在此基础上,明确了肌肉细胞生长规律以及诱导分化后符合运动属性特征的超微结构。Somatic cell cultured in vitro is the most advantageous tool to study growth and development of muscle tissue.However,the in vitro culture system of muscle cells for sheep has not been succeeded so far,and there are little reports on growth feature of muscle cells and ultrastructure after differentiation.In this study,muscle cells of Luxi blackhead sheep were cultured by enzymatic digestion,and the myogenicity was identified by laser confocal fluorescence microscopy.The growth curve was drawn to research the cell growth law.The differentiation medium was adopted to induce myogenesis,and the ultrastructure was observed through scanning electron microscope and transmission electron microscope.The results were as follows.The culture in vitro of muscle cells from Luxi blackhead sheep had been established and the cell purity was more than 95%.The growth of muscle cells experienced incubation period,exponential growth period,plateau period and decayed period.The plateau period was relatively longer,during which,muscle cells differentiated spontaneously.After inducted differentiation,cell surface protruded silk feet,and plasma membrane was distributed with many voids;mitochondria were very rich,and inner cristae was developed and dense;a large number of rough endoplasmic reticulum existed,and the surface ribosomes were clearly visible;a large amount of lipids and glycogen were scattered in the cytoplasm;myofibril bundles were dense and arranged regularly.In conclusion,the culture system of muscle cells from Luxi blackhead sheep had been successfully established.On this basis,we clarified the growth fecture of muscle cells and the ultrastructure with distinctive characteristics after induced differentiation.

关 键 词：肌肉细胞 诱导分化 超微结构 鲁西黑头羊 

分 类 号：S826.9[农业科学—畜牧学]