原代藏獒细胞培养与生物学特性研究  

Primary Culture and Biological Characteristics of Tibetan Mastiff Kidney Cells

作　　者：刘文凯 田玲 乔自林[1,3] 李铀 王家敏[1,3] 王明明[1,2] LIU Wen-kai;TIAN Ling;QIAO Zi-lin;LI You;WANG Jia-min;WANG Ming-ming(Gansu Tech Innovation Center of Animal Cell,Biomedical Research Center,Northwest Minzu University,Lanzhou 730030,PRC;Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission,Biomedical Research Center,Northwest Minzu University,Lanzhou730030,PRC;College of Life Science and Engineering,Northwest Minzu University,Lanzhou 730030,PRC)

机构地区：[1]西北民族大学生物医学研究中心甘肃省动物细胞技术创新中心,甘肃兰州730030 [2]西北民族大学生命科学工程学院,甘肃兰州730030 [3]西北民族大学生物医学研究中心生物工程与技术国家民委重点实验室,甘肃兰州730030

出　　处：《湖南农业科学》2021年第11期1-4,共4页Hunan Agricultural Sciences

基　　金：西北民族大学中央高校基本科研业务费专项资金(31920210002,31920200004);甘肃省科技计划项目(21JR1RA222,18JR2JA004)。

摘　　要：为研究藏獒体细胞的生物学特性,保存藏獒遗传资源,以实验室前期自主分离的藏獒肾脏细胞为材料,探讨了其体外培养的形态特征、传代次数、冻存复苏、生长特性、细胞核型以及G418最低致死浓度。结果表明:藏獒肾脏细胞体外培养时形态均为成纤维细胞,形状梭形,呈放射状与旋涡状样式生长;细胞冻存后复苏活率为91%,且复苏后形态与长势均呈现良好状态;F_(1)代至F_(5)代,细胞增殖较快,F_(5)代至F_(8)代,细胞增殖减弱甚至停滞,细胞呈多边形状态,出现细胞脱落现象,最终于F_(8)代无法继续生长;生长曲定结果显示,24孔板内细胞数目最高值达到4.06×10^(5)个/mL,最短倍增时间为27 h;藏獒核型2n=78;藏獒肾脏细胞的G418筛选最低致死浓度为500μg/mL。In order to study the biological characteristics of Tibetan mastiff somatic cells and preserve the genetic resources of Tibetan mastiff,this study conducted the primary culture of the pre-isolated kidney cells of Tibetan mastiff,and revealed the morphological characteristics,generation numbers,cryopreservation and cell thawing,growth characteristics,karyotype,and G418 minimum lethal concentration in vitro culture.The results showed that the Tibetan mastiff kidney cells cultured in vitro were spindle-shaped fibroblast in radial or swirl growth.With a resuscitation rate of 91%,the survial renal cells grew well in good morphology.From F_(1) generation to F_(5) generation,the cells proliferated rapidly.From F_(5) generation to F_(8) generation,the cell proliferation weakened or even stagnated,and the cells exhibited polygonal shape,occurring the phenomenon of exfoliation,finally could not continue growing in F_(8) generation.According to the measured result of growth curve,the maximum cell number on the 24-well microplate reached 4.06×10^(5) cells/mL and the shortest cell population doubling time was 27 h.The karyotype of Tibetan mastiff is 2n=78.The lowest lethal concentration of G418 screened by Tibetan mastiff kidney cells is 500μg/ml.

关 键 词：藏獒 细胞培养 G418 

分 类 号：Q813.11[生物学—生物工程]