《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (2): 586-593.doi: 10.16431/j.cnki.1671-7236.2017.02.039

• 基础兽医 • 上一篇    下一篇

现代商品仔猪传代肾细胞株的培育与应用研究

胡鸿惠, 吴静波, 黄健强, 南文金, 于文涛   

  1. 韶关学院动物疫病实验室, 韶关 512005
  • 收稿日期:2016-07-18 出版日期:2017-02-20 发布日期:2017-02-25
  • 通讯作者: 胡鸿惠 E-mail:beijinggd@163.com
  • 作者简介:胡鸿惠(1982-), 女, 湖南郴州人, 硕士, 助理研究员, 研究方向:动物生产与疾病
  • 基金资助:

    广东省协同创新平台“粤北生猪生产及疫病防控协同创新发展中心”(YJK-2014-52-16);广东省现代农业产业技术体系建设专项资金;韶关市科技计划项目(2013CX/NO6、韶科[2015]72号);广东省科技计划项目(2016A040402038);韶关学院科研项目(SY2016KJ24)

Study on Cultivation Research and Preliminary Application of Modern Commercial Piglet Kidney Cell Strain

HU Hong-hui, WU Jing-bo, HUANG Jian-qiang, NAN Wen-jin, YU Wen-tao   

  1. Animal Disease Laboratory, Shaoguan University, Shaoguan 512005, China
  • Received:2016-07-18 Online:2017-02-20 Published:2017-02-25

摘要:

细胞系是动物病毒分离培养的重要载体。本研究以现代商品化仔猪肾脏为原始材料,拟培育新的细胞系用于动物病毒的分离和培养。利用胰酶消化法和差速贴壁相结合方法,分离纯化仔猪肾上皮细胞,并在体外进行传代培养和筛选。结果显示,试验成功得到一株可以连续传代的细胞株,命名为SDPK-D,且已在体外连续传代90代。SDPK-D细胞株F33和F83代倍增时间分别为40.9和32.7 h,细胞活率分别为97.55%和98.86%,8 h细胞贴壁率分别为91.67%和97.06%。在该细胞株接种猪伪狂犬病病毒(PRV)、猪流感病毒(SIV)、猪流行性腹泻病毒(PEDV)阳性病料均出现明显的细胞病变。本研究首次针对现代商品猪培育出一株可以在体外连续传代的细胞株,并对多种动物病毒敏感,为相关动物病毒的分离培养提供了新的细胞系选择。

关键词: 仔猪肾细胞; 原代培养; 传代培养; 细胞株

Abstract:

Cell line was an important carrier for the isolation and culture of animal viruses.In the research, modern commercial piglet kidney was taken as the raw material in order to cultivate a new cell line for isolating and culturing animal viruses. By trypsin digestion and differential velocity adherent combination method,porcine kidney epithelial cells were separated and purified,and then continuously subcultured in vitro. The results showed that a new cell strain named SDPK-D had been serially passaged for 90 generations.The doubling time of SDPK-D cell strain F33 and F83 were 40.9 and 32.7 h,respectively;While the cell viability were 97.55% and 98.86%,the cell adherent rate at the 8 h were 91.67% and 97.06%,respectively. Obvious cytopathic effect appeared after inoculating with PRV,SIV and PEDV positive samples. For the first time, a new cell strain from modern pig named SDPK-D was successfully cultivated,which was sensitive for several animal viruses.It could provid a new selection for the isolation and culture of virus.

Key words: piglet kidney cell; primary culture; subculture; cell strain

中图分类号: