猪肌肉卫星细胞的分离培养及生物学特性鉴定
Isolation and characterization of porcine muscle satellite cells
摘要 以猪胎儿为材料,采用胶原酶消化法或组织块法培养胎儿背部最长肌获得了肌肉卫星细胞,该细胞体外可以传到9代以上。培养的细胞多呈纺锤体型和梭型,具有明显的方向性,呈典型的长轴平行排列。流式细胞仪分析结果显示,该细胞呈CD29、CD166、CD45、CD44阳性,CD71、CD34阴性。RT-PCR检测发现其表达Desmin、C-Myc、Nanog、Pcna、Oct4、Klf4,弱表达Myog,不表达Sox2、MyoD。免疫组化染色发现其表达Desmin等肌肉细胞的特异性标记,同时表达Nanog、Pcna等多能性细胞标记。本试验建立了一种简便高效的猪肌肉卫星细胞体外分离和培养方法,得到的细胞具有肌肉卫星细胞的典型生物学特性,同时表达间质干细胞和多能性干细胞的部分标记。
Abstract:
The muscle satellite cells were got from culturing the fetal longissimus muscle with the methods of enzyme digestion and tissue culture.The cells were cultured more than 9 passages,which were mostly spindle-type and spindle-like,and they possessed visible direction and parallel typically.These cells were postive for CD29,CD166,CD44,CD45 and negative for CD71 and CD34 analysed by flow cytometry.RT-PCR analysis showed that the cells were postive for Desmin,C-Myc,Nanog,Pcna,Oct4 and Klf4,weakly positive for Myog and negative for Sox2 and MyoD.By immunohistochemistry it was discovered that Myog,Desmin,Nanog and Pcna were expressed in the cells.Results showed that a simple and efficient system was established for isolation and culture of porcine muscle satellite cells,and the cells possessed typical characteristics of muscle statellite cells,also the cells shared features of mesenchymal stem cells and pluripotent stem cells.
作者单位: 西北农林科技大学动物医学院陕西省干细胞工程技术研究中心农业部动物生殖发育与胚胎工程重点开放实验室陕西省农业分子生物学重点实验室,陕西杨凌,712100
刊 名 中国兽医学报 ISTICPKU
年,卷(期): 2011, 31(10)
分类号: S814.8
机标分类号: R5 R73
基金项目: 国家自然科学基金资助项目,教育部新世纪优秀人才支持计划资助项目