【摘要】 为了建立简便、高效的仔猪小肠平滑肌细胞体外培养方法,为相关研究提供试验材料,采用组织贴块法和酶消化法进行原代培养,胰酶消化传代,并应用倒置显微镜和SABC试剂盒对细胞进行形态学和免疫组化鉴定。结果采用组织块贴块法培养的仔猪肠平滑肌细胞有90%的组织块接种存活,初次传代后的平滑肌细胞纯度达95%以上。免疫组化染色显示胞浆内α平滑肌肌动蛋白阳性表达。而酶消化法培养的仔猪肠平滑肌细胞,生长缓慢甚至死亡。表明组织块贴块法是为简便、可靠,短期内可获大量高纯度的仔猪小肠平滑肌细胞的原代培养方法。更多还原

【Abstract】 In order to establish a simple and effective method of culturing piglet’s intestinal smooth muscle cells in vitro,and to provide test material for the related studies,tissue-piece inoculation and collagenase digestion were adopted to culture the primary piglet’s intestinal smooth muscle cells,respectively,and then rypsin digestion was used to transfer them.The cultured cells were identified by morphology and immunohistochemistry with inverted microscope and SABC kit.The result showed that about 90%inoculated tissue pieces survived through tissue-piece inoculation and the purity of the piglet’s intestinal smooth muscle cells by the first passage was more than 95%.And these cells were positive with the immunohistochemically staining against mouse SM-α-actindemonstrated by specific mAb.At the same time,the cells cultured by collagenase digestion grew very slowly,or even died.Our results indicate that tissue-piece inoculation is a simple and reliable method to obtain highly purified pigler’ s intestinal smooth muscle cells in short term.更多还原