转瓶培养与生物反应器微载体培养重组腺病毒的比较

发布时间：2018-04-20 02:31

本文选题：转瓶 + 生物反应器　；参考：《中国病原生物学杂志》2016年01期

【摘要】：目的分别用3L转瓶和7L生物反应器微载体系统培养人胚胎肾细胞(HEK-293细胞)并接种H3型流感重组腺病毒,比较两种方法培养细胞及重组腺病毒的效果。方法分别用3L转瓶和7L生物反应器培养HEK-293细胞。转瓶每24h取样,生物反应器每12h取样,记录细胞总数。待细胞长满表面85%~90%时接种流感重组腺病毒,完全病变后收取病毒液并测定病毒滴度。结果转瓶培养HEK-293细胞3d,细胞增殖增加1.85~2.45倍,为(3.71~4.90)×107个[(2.95~3.90)×104 cells/cm2],生物反应器培养3d,细胞增长4.72~5.00倍,为(132.15~140.09)×107个[(24.88~28.72)×104cells/cm2]。转瓶培养重组腺病毒滴度为6.85TCID50/ml,生物反应器微载体系统培养腺病毒滴度为7.200TCID50/ml。结论生物反应器微载体系统培养流感重组腺病毒数量和滴度均高于转瓶培养,不但提高了疫苗的产量,还减少了细胞分泌物的残留,增强了疫苗接种的安全性。
[Abstract]:Objective to culture human embryonic kidney cells (HEK-293 cells) with 3L flask and 7L bioreactor respectively and inoculate H3 influenza recombinant adenovirus. Methods HEK-293 cells were cultured in 3 L flask and 7 L bioreactor respectively. The total number of cells was recorded in bioreactor and bioreactor every 24 hours. After inoculating influenza recombinant adenovirus, the virus solution was collected and the titer of the virus was measured. Results the proliferation of HEK-293 cells was increased by 1.85 ~ 2.45 times, which was 3.71 ~ 4.90 脳 10 ~ (7) 脳 10 ~ 4 cells/cm2 (2.95 ~ 3.90) 脳 10 ~ 4 cells/cm2 for 3 days. The cell growth was 4.72 ~ 5.00 times in a bioreactor for 3 days, which was 132.15 ~ 140.09) 脳 10 ~ (7) 脳 10 ~ (7) [24.8828.72 脳 104cells/cm2]. The titer of recombinant adenovirus in flask culture was 6.85 TCID 50 / ml, and the titer of adenovirus in bioreactor microcarrier system was 7.200 TCID 50 / ml. Conclusion the amount and titer of recombinant adenovirus in bioreactor microcarrier system are higher than those in flask culture, which not only increases the production of vaccine, but also reduces the residue of cell secretion and enhances the safety of vaccination.
【作者单位】：吉林农业大学动物科学技术学院;军事医学科学院军事兽医研究所;吉林大学动物医学学院;延边大学医学院;
【基金】：国家863项目(No.2012AA02A403) 国际合作交流项目(No.2014DFA30290) 吉林省自然科学基金项目(No.20130101093JC)
【分类号】：R511.7;R450

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