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| 欧拉羊胚肾细胞的分离培养和鉴定 | |
| 引用本文： | 乔自林,李明生,冯若飞,令世鑫,王家敏,沈武玲,冯玉萍,柏家林,马忠仁.欧拉羊胚肾细胞的分离培养和鉴定[J].安徽农业科学,2012,40(2):850-851,980. |
| 作者姓名： | 乔自林 李明生 冯若飞 令世鑫 王家敏 沈武玲 冯玉萍 柏家林 马忠仁 |
| 作者单位： | 1. 甘肃省动物细胞工程技术研究中心,甘肃兰州730030;西北民族大学生命科学与工程学院,甘肃兰州730030 2. 甘肃省动物细胞工程技术研究中心,甘肃兰州,730030 3. 西北民族大学生命科学与工程学院,甘肃兰州,730030 |
| 摘 要： | 目的]分离培养并鉴定欧拉羊胚肾细胞,为该品种基因组文库构建和遗传多样性研究提供生物学材料。方法]用胰蛋白酶热消化法分离并培养欧拉羊胚肾原代细胞,通过差速消化和差速贴壁法纯化成纤维细胞,扩大培养至第3代用液氮保存,细胞复苏后进行活力、形态、生长曲线、微生物污染检测和连续传代培养试验。结果]欧拉羊胚肾细胞呈成纤维型,复苏活力90%以上,生长良好,生长曲线呈"S",最大增殖浓度为4.61×105/ml,倍增时间26 h;细菌、真菌、病毒、支原体检测呈阴性,连续传代培养在10代内生长正常。结论]欧拉羊胚肾细胞分离培养成功,使这一重要种质资源在细胞水平上得以保存。 |
| 关 键 词： | 欧拉羊 胚肾细胞 培养 鉴定 |
Separation, Culture and Identification of Embryo Kidney Cells in Oula Sheep |
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| Institution: | QIAO Zi-lin et al(Gansu Research Center for Animal Cell Engineering,Lanzhou,Gansu 730030) |
| Abstract: | Objective] The study aimed to separate,culture and identify the embryo kidney cells in Oula sheep and provide the biological material for the establishment of the genome library and genetic diversity of this variety.Method] The primary cells of the embryo kidney in Oula sheep were separated and cultured with the trypsin digestion method,purified into the fibroblasts through the differential digestion and differential adherence isolation methods and preserved with the liquid nitrogen when expanding the culture to the third generation.The observations on the vitality,morphology and dynamic growth,the detection on the microbial contamination and the test on the continuous subculture were carried out after the recovery of the cells.Result] The cells of embryo kidney in Oula sheep were fibroblast and the vitality of the recovery cell was over 90%;the cells grown well,with the growth curve showing "S" type,the maximum proliferation concn.of 4.61×105/ml and the population doubling time(PDT) of 26 h;the detection on the bacteria,fungi,virus and mycoplasma showed all negative and the cells grown normally after being subcultured within 10 generations.Conclusion] The cells of the embryo kidney in Oula sheep were separated and cultured successfully,which made the important genetic resource to be preserved in the cell level. |
| Keywords: | Oula sheep Embryo kidney cell Culture Identify |
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