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当前位置: 首页 > 文章 > 犬瘟热病毒Vero细胞悬浮产业化培养工艺研究 现代畜牧兽医 2022 (1) 57-60
Position: Home > Articles > Study on the suspended industrial culture technology of dog distemper virus Vero cells Modern Journal of Animal Husbandry and Veterinary Medicine 2022 (1) 57-60

犬瘟热病毒Vero细胞悬浮产业化培养工艺研究

作  者：
郑新勇;闫福海
单  位：
天津瑞普生物技术股份有限公司空港经济区分公司;山东省鲁茂生态科技有限公司
关键词：
犬瘟热;Vero细胞;反应器;悬浮培养
摘  要：
为克服犬瘟热疫苗现有生产工艺的缺陷,试验采用10 g/L Cytodex-1型微载体,按每个微载体15～20个细胞的细胞接种量接种至微载体培养Vero细胞,细胞培养液为10%NBS的DMEM培养液。结果显示,当细胞密度达到8×106CFU/mL时接种犬瘟热病毒,最佳培养时间30 h,接毒剂量按照MOI为0.1接种犬瘟热病毒液;当细胞病变达到50%时,病毒感染细胞时间为30 h,收获毒液。按照上述摸索生产工艺参数,收获的犬瘟热病毒液的病毒液滴度每病毒含量≥108.5TCID50/0.1 mL。将收获的病毒液冻存及下游相关的灭活处理,作为制备犬瘟热疫苗的抗原。研究表明,试验大幅度提升犬瘟热病毒培养量,效价批间差异性均一,实现了产业化反应器悬浮培养代替细胞工厂的技术路线。
译  名：
Study on the suspended industrial culture technology of dog distemper virus Vero cells
作  者：
Zheng Xianyong;Yan Fuhai;Tianjin Ruipu Biotechnology Co.Ltd.,Airport Economic Branch Company;Shandong Lumao Ecological Technology Co.Ltd.;
关键词：
Canine distemper;;Vero cells;;Reactor;;Suspension culture
摘  要：
In order to overcome the shortcomings of the current production process of canine distemper vaccine, the experiment used 10 g/L Cytodex-1 type microcarriers, and inoculated the microcarriers to culture Vero cells according to the cell inoculation amount of 15～20 cells per microcarrier. The cell culture medium was 10% NBS DMEM medium. When the cell density reached 8×10~6 CFU/mL for inoculation of canine distemper virus, the best culture time was 30 h, and the dose of inoculation was based on MOI of 0.1 for inoculation of canine distemper virus solution. When the cytopathic effect reached 50%, the virus infected the cells for 30 hours, and the venom was harvested. According to the above-mentioned production process parameters, the virus titer of the harvested canine distemper virus liquid was ≥108.5 TCID50/0.1 mL, and the harvested virus liquid was frozen and stored and downstream related inactivation treatment was used as the preparation of the canine distemper vaccine antigen. The study indicates that the experiment greatly increase the amount of canine distemper virus cultured, and the titers has uniform differences between batches, realizing the technical route of industrial reactor suspension culture instead of cell factories.

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