鲫脑组织细胞与鲤疱疹病毒Ⅱ型的微载体规模化培养工艺
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- 英文论文题名：Technologies for large-scale cultivation of gibel carp brain cells and cyprinid herpesvirus 2 by microcarrier
- 论文作者：王维玲 ; 马杰 ; 周勇 ; 范玉顶 ; 刘文枝 ; 曾令兵
- 英文论文作者：WANG Weiling ; MA Jie ; ZHOU Yong ; FAN Yuding ; LIU Wenzhi ; ZENG Lingbing ; College of Fisheries and Life Science ; Shanghai Ocean University ; Yangtze River Research Institute ; Chinese Academy of Fishery Sciences
- 年：2016
- 作者机构：上海海洋大学水产与生命科学学院;中国水产科学研究院长江水产研究所;
- 论文关键词：鲫脑组织细胞系(GiCB) ; 鲤疱疹病毒2型(CyHV-2) ; 微载体 ; 规模化培养 ; 工艺
- 英文论文关键词：GiCB ; cyprinid herpesvirus 2(CyHV-2) ; microcarrier ; large-scale culture
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：S948
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：508k
- 原文格式：D
- 会议级别：全国
摘要
本文研究了在Cephodex微载体悬浮培养系统中规模化培养鲫脑组织细胞(GiCB)和鲤疱疹病毒Ⅱ型(Cyprinid herpesvirus Ⅱ,CyHV-2)的工艺。结果表明,微载体Cephodex适合GiCB细胞的贴壁培养,细胞贴壁期培养基中血清浓度为10%,微载体浓度为6g/L;细胞初始接种密度为2.5×10~5 cells/ml时,以转速35r/min,每静置30min搅拌2min的间歇搅拌方式贴壁率最佳,8h后贴壁率可达90%以上;增殖期以45r/min的连续搅拌速度可获得最佳的细胞生长效能。采用优化的工艺条件,以感染复数为0.2的CyHV-2病毒接种规模化培养的GiCB细胞5d后,出现典型的细胞病变效应,病毒滴度(TCID50/ml)可达106.5。本项研究为鲫造血器官坏死症疫苗的规模化制备技术研究奠定了前期基础。
In this paper the technologies for the large-scale cultivation of gibel carp brain cells and CyHV-2 by Cephodex microcarrier in suspension system were studied. The results showed that the Cephodex is a kind of microcarrier suitable for the growth of anchorage-dependent GiCB cells.The optimal serum concentration for cells attaching to the Cephodex microcarrier was 10%,the microcarrier concentration was 6g/L and the density of cells inoculated was 2.5×10~5cells/ml.With an intermittent agitation for 2 min at 35 rpm after 30 min stilling culture, the attachment efficacy reached 90% after 8h cultivation While in growing period, The optimal agitation speed was 40 rpm for continuous stirring. After infection of the GiCB cells on Cephodex microcarrier with CyHV-2 at a multiplicity of infection(MOI) of 0.2, the typical cytopathic effect(CPE) appeared after 5 days post-infection and the virus titer(TCID50/ml) reached 106.5.
In this paper the technologies for the large-scale cultivation of gibel carp brain cells and CyHV-2 by Cephodex microcarrier in suspension system were studied. The results showed that the Cephodex is a kind of microcarrier suitable for the growth of anchorage-dependent GiCB cells.The optimal serum concentration for cells attaching to the Cephodex microcarrier was 10%,the microcarrier concentration was 6g/L and the density of cells inoculated was 2.5×10~5cells/ml.With an intermittent agitation for 2 min at 35 rpm after 30 min stilling culture, the attachment efficacy reached 90% after 8h cultivation While in growing period, The optimal agitation speed was 40 rpm for continuous stirring. After infection of the GiCB cells on Cephodex microcarrier with CyHV-2 at a multiplicity of infection(MOI) of 0.2, the typical cytopathic effect(CPE) appeared after 5 days post-infection and the virus titer(TCID50/ml) reached 106.5.
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