鸭瘟病毒在鸭胚脐带间充质干细胞上生物学特性的研究

发布时间：2018-07-11 10:22

本文选题：鸭瘟病毒 + 鸭胚脐带间充质干细胞　；参考：《中国兽医科学》2017年09期

【摘要】：为了解鸭瘟病毒(DPV)在鸭胚脐带间充质干细胞上的适应性和增殖情况。本试验将DPV接种鸭胚脐带间充质干细胞,连续传代至第10代,通过CPE观察、病毒粒子电镜观察、核酸检测、间接免疫荧光检测等分析其生物学特性,并测定了连续10代的病毒滴度(TCID_(50))。结果显示,第一代DPV即在鸭胚脐带间充质干细胞上产生了明显的CPE,并且在连续传代的过程中CPE能够稳定出现。初步提纯的病毒悬浮液在电镜下可见到结构清晰的子代病毒。F1~F10代DPV均能扩增出大小为416 bp的UL6基因片段。间接免疫荧光又进一步证实DPV的阳性血清能够对染毒病灶产生特异性荧光染色。连续测定了10代的病毒滴度(TCID_(50))均不低于10~(-6.43),且在DEUCMSC上测定的DPV TCID_(50)明显高于DEF的。说明DPV能在鸭胚脐带间充质干细胞上稳定增殖。
[Abstract]:To investigate the adaptability and proliferation of duck plague virus (DPV) on duck umbilical cord mesenchymal stem cells. In this experiment, DPV was inoculated with duck umbilical cord mesenchymal stem cells. The biological characteristics of DPV were analyzed by CPE observation, electron microscope observation of virus particles, nucleic acid detection, indirect immunofluorescence detection and so on. The virus titer (TCID _ (50).) of 10 consecutive generations was determined. The results showed that the first generation of DPV produced significant CPE on the umbilical cord mesenchymal stem cells of duck embryo, and CPE appeared stably during successive passage. The purified virus suspensions could amplify the UL6 gene fragment of 416 BP in DPV of F1 F10 generation with clear structure under electron microscope. Indirect immunofluorescence further confirmed that the positive serum of DPV could produce specific fluorescent staining on the infected lesions. The virus titers (TCID50) of 10 generations were measured continuously, and the DPV TCID50 on DEUCMSC was significantly higher than that of DEF. It indicated that DPV could proliferate steadily on the mesenchymal stem cells of duck embryo umbilical cord.
【作者单位】：东北农业大学生命科学学院;中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室黑龙江省实验动物与比较医学重点实验室;
【基金】：国家科技支撑计划项目(2015BAI07B02-02) 中国农业科学院基本科研业务费专项(Y2016PT41) 黑龙江省自然科学基金重点项目(ZD2016006) 中央级公益性科研院所基本科研业务费专项(1610302016013,1610302017013)
【分类号】：S852.65

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