F81细胞无血清悬浮驯化后培养猫泛白细胞减少症病毒的研究  

Serum-free Suspension Domestication of F81 Cell Line for Culture of Feline Panleukopenia Virus

作　　者：张久鑫 邵宝顺 朱辉 贾浩苒 梁琳[2] 汤新明 崔尚金[2] 梁瑞英 王龙涛[1] ZHANG Jiu-xin;SHAO Bao-shun;ZHU Hui;JIA Hao-ran;LIANG Lin;TANG Xin-ming;CUI Shang-jin;LIANG Rui-ying;WANG Long-tao(College of Animal Science and Technology,Jilin Agricultural University,Changchun,Jilin,130118,China;Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing,100089,China;Guangdong Huasheng Biotechnology Co.,Ltd.,Guangzhou,Guangdong,511300,China)

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118 [2]中国农业科学院北京畜牧兽医研究所,北京100089 [3]广东省华晟生物技术有限公司,广东广州511300

出　　处：《动物医学进展》2023年第1期48-53,共6页Progress In Veterinary Medicine

基　　金：中央科研院所基本科研业务经费项目(2020-YWF-YTS-10);中国农业科学院创新工程计划项目(ASTIP-IAS15)。

摘　　要：为解决国内猫泛白细胞减少症生物制品短缺的问题,筛选出适用于猫泛白细胞减少症病毒增殖的猫肾细胞系(F81),通过对贴壁F81进行降血清悬浮驯化培养,得到了无血清全悬浮培养F81细胞系。研究建立的F81细胞系在初始传代细胞密度为1×10^(6)cells/mL时,培养48 h后细胞密度可达到7.02×10^(6)cells/mL。对培养猫泛白细胞减少症病毒工艺进行摸索,确定最佳接毒工艺为同步接毒法,初始接毒细胞密度为1×10^(6)cells/mL、MOI为3.5×10^(-2),培养72 h后收毒,病毒滴度可达到10^(7.25)TCID_(50)/0.1 mL。该方法使用同源细胞系且未添加动物血清,为猫泛白细胞减少症生物制品生产降低了下游生产纯化难度,同时降低了生产成本,可显著提高产品核心竞争力。研究结果可为其他猫用生物制品相关研究提供参考。In order to solve the shortage of feline panleukopenia biological products in China,a feline kidney cell line(F81)suitable for the propagation of feline panleukopenia virus was screened out.The serum-free F81 cell line was obtained by domesticating the adhesion F81 with serum-lowering suspension culture.The cell density of the F81 cell line established in this study reached 7.02×10^(6) cells/mL after 48 hours of culture when the cell density of the initial passage was 1×10~6 cells/mL.To explore the technology of feline panleukopenia virus culture,the best technology was determined as:synchronous inoculation method,the initial inoculation cell density was 1×10^(6) cells/mL,the MOI was 3.5×10^(-2),after 72 hours of culture,the virus titer could reach 10^(7.25)TCID_(50)/0.1 mL.This method uses homologous cell lines and does not add animal serum,which reduces the difficulty of downstream production and purification for the production of feline panleukopenia biological products,and also reduces the cost,which can significantly improve the core competitiveness of the product.The results of this study can provide reference for other biological products for cats.

关 键 词：F81细胞 细胞驯化 无血清悬浮培养 猫泛白细胞减少症病毒 

分 类 号：S852.659.2[农业科学—基础兽医学] S858.293[农业科学—兽医学]