Ⅰ型禽腺病毒AV208株在鸡肝癌细胞中增殖规律的研究  

Growth pattern of fowl adenovirus typeⅠ-AV208 strain on chicken hepatocellular carcinoma cell line,LMH

作　　者：赵莉[1,2] 周洁 高诚 胡建华 

机构地区：[1]上海实验动物研究中心,上海201203 [2]扬州大学兽医学院,江苏扬州225009

出　　处：《微生物学通报》2012年第8期1120-1126,共7页Microbiology China

基　　金：上海市科研计划项目(No.09140901400)

摘　　要：【目的】摸索鸡肝癌细胞系(LMH)培养条件,并采用LMH细胞系对Ⅰ型禽腺病毒AV208株(FAVⅠ-AV208)进行增殖规律的研究。【方法】细胞以不同血清浓度培养并以不同接种比例传代。观察最佳感染复数下的细胞病变和病毒增殖情况,并研究病毒接种物浓度与蚀斑形成数量之间的关系。【结果】LMH细胞系的最适培养条件为,以10%FBS的培养基以1:5比例传代培养。FAVⅠ-AV208毒株可以在LMH细胞中良好增殖并达到较高的滴度(107.5TCID50/0.1 mL)。在最佳感染复数(MOI)为0.01时,72 h细胞即出现明显的细胞病变(CPE),特征为细胞变大变圆,集聚成不规则的葡萄串状。病毒接种物浓度与蚀斑形成数量间呈线性相关。【结论】LMH细胞是FAVⅠ较合适的病毒培养系统,为研制禽腺病毒重组疫苗提供了有利工具。[Objective] To explore culture conditions for chicken hepatocellular carcinoma cellline （LMH） and growth pattern of fowl adenovirns type Ⅰ-AV208 strain （FAV Ⅰ-AV208） on LMH cells. IMethods] Cells were cultured with different concentration of serum and propagated at dif- ferent subcultivation ratios. Cytopathic effect and growth pattern ofFAV Ⅰ- AV208 on LMH cells were studied at an optimal MOI. Relation between virus concentration and plaque number was discussed by a standard plaque assay. [Results] Culturing with 10% FBS is suitable for LMH cells and a subcultivation ratio of 1：5 is recommended. FAV Ⅰ- AV208 could be propagated efficiently in LMH cells and high virus titer up to 107.5 TCIDs0/0.1 mL was reached. Obvious CPE was observed after 72 h post- infection at an MOI of 0.01. The nomal cobblestone- like pattern cells grew into larger and round, they aggregated into irregularly botryoidal appearance and finally disintergrated. A linear relation was proved between virus concentration and plaque number. [Conclusion] LMH cell line is a suitable virus culture system and provides an excel- lent tool for constrution of recombinant fowl adenovirus type Ⅰ .

关 键 词：Ⅰ型禽腺病毒AV208株 鸡肝癌细胞 细胞病变 增殖规律 

分 类 号：S852.65[农业科学—基础兽医学]