%A LU Bingxia, TAO Minghua, ZHAO Shuo, QIN Yibin, LIU Lei, CHEN Zhongwei, HE Ying, DUAN Qunpeng, LIANG Jiaxing, ZHOU Yingning, LI Bin, JIANG Dongfu, YANG Siyi, ZHAO Wu %T Optimization of Large-scale Rotary Bottle Culture Conditions for Porcine Epidemic Diarrhea Virus Mutant CH/GX/750A/2015 %0 Journal Article %D 2020 %J China Animal Husbandry and Veterinary Medicine %R 10.16431/j.cnki.1671-7236.2020.05.025 %P 1506-1513 %V 47 %N 5 %U {https://www.chvm.net/CN/abstract/article_38822.shtml} %8 2020-05-20 %X The purpose of this study was to determine the optimal conditions for the rotation culture of CH/GX/750A/2015 strain of porcine epidemic diarrhea virus (PEDV) on Vero cells,and provide technical support for the mass production of high-efficiency vaccine.The virus was cultured on Vero cells in a 10 L spinner flask.The conditions of trypsin concentrations(0,2,4,6,8,10 μg/mL),cell density(40%,60%,80%,100% and 200%),virus inoculation dose (MOI=1,0.1,0.01 and 0.001),virus adsorption time (0,30,60,90,120 and 240 min),virus harvest time (12,24,36,48,60,72 h after virus inoculation),culture temperature after virus inoculation(35,36,37 and 38 ℃) and spinner speed(6,8,10,12,14 and 16 r/h) were optimized.The results showed that the best culture condition of PEDV CH/GX/750A/2015 strain in 10 L transfer flask was to inoculate the virus when the cell was just covered with a single layer,the dose of virus was MOI=0.01,the concentration of trypsin in the culture medium after virus inoculation was 6 μg/mL,without adsorption,the culture temperature after virus inoculation was 37 ℃,the best spinner speed 12 r/h,and the best virus harvest time was 48 h after virus inoculation.After optimization,the virus titer can reach 106.50 TCID50/0.1 mL.This study provided a reference for the large-scale culture of PEDV mutant strain,and laid a foundation for the development of high-efficiency vaccine of PEDV mutant strain.