鸡IBDV在微载体微型反应器DF-1细胞系上繁殖特性研究  

Research on the breeding characters of IBDV on DF-1 cell at micro carrier in tubular miniature reactor

作　　者：杨霞[1] 周欣[1] 陈陆[1] 王永生[1] 赵军[1] 王川庆[1] 王泽霖[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002

出　　处：《中国兽医学报》2013年第10期1519-1526,共8页Chinese Journal of Veterinary Science

基　　金：国家农业科技成果转化基金资助项目(2011GB2D000007);国家自然科学基金资助项目(30972187)

摘　　要：对IBDV在微型生物反应器转管DF-1细胞系上繁殖特性进行了较为系统的研究，转管中接种DF-1细胞3×10’个左右，培养6d细胞增至2．4～2．5×10^8个，确定培养6d为最佳接毒时间；病毒的最佳接种剂量为0．78TCID50／细胞（即0．78MOI），最佳收毒时间为接毒后的28h；上清和全悬液毒价呈平行关系，但后者更高，可达10^9.5TClD5.0／mL以上。细胞数增长与糖耗间呈明显平行关系，在细胞生长期内（6d）平均每个细胞耗糖量为6．5×10μg／24h，可根据糖耗量的多少推测细胞生长状态和数量；接毒后糖耗速率的高峰要先于病毒滴度高峰，在糖耗高峰出现后下降至趋于零的瞬间收获，可获得较高毒价的病毒液。在毒价高峰时收获1／2毒液后9h，收获1／3毒液后18h可再次获得最高毒价的毒液，这可能与高密度培养中及时补充新的培养液有关。本试验为IBDVHQ株在生物反应器上大规模培养提供了参考依据。This article comprehensively researched on the breeding characters of IBDV on DF-1 cell at micro carrier in tubular miniature reactor. Inoculated in tubular about with 3 X 10^7 or so,DF-1 cells increased up 2.4--2.5 X 10^8after inoculated 6 days,which was determined as the best inocula tion time for IBDV; optimal inoculation dose of IBDV was 0.78 TCIDs0/eell （0.78 MOI） ,the best time of collection was at 28 h after virus inoculation;there was a parallel correlation of virus titers between supernatant and cell suspension, but the latter, could achieve higher than 10^9.5 TCIDs0/mL. The number of cells growth and sugar consumption showed obvious parallel correlation, the average sugar consumption of each cell in cell growth period （6 days ） was up to 6.5X 10-10 g/24 h,according to sugar consumption rate we could calculated the number and estimated growth state of DF-1 cell;the peak time of sugar consumption was prior to the peak of virus titers after inoculation,when the sugar consumption fall from pesks to zero instantaneity,a higher virus titers could be obtain. It could again obtain the higher virus titers at 9 h after collecting 1/2 of virus solution at peak time,and at 18 h after harvesting 1/3 ,which may be associated with that after inoculated cells timely obtained nutrition from new liquid in high density culture. This experiment provides reference for IBDV HQ cultivating in DF-1 cell on micro carrier at tubular miniature reactor at a certain extent.

关 键 词：传染性法氏囊病毒 DF-1细胞 转管微载体微型反应器 病毒感染复数 糖耗 

分 类 号：S852.65[农业科学—基础兽医学]