Ⅰ型禽腺病毒AV208株在鸡肝癌细胞中增殖规律的研究

摘要

[Objective]To explore culture conditions for chicken hepatocellular carcinoma cell line (LMH) and growth pattern of fowl adenovirus type Ⅰ-AV208 strain (FAV Ⅰ -AV208) on LMH cells.[Methods] Cells were cultured with different concentration of serum and propagated at different subcultivation ratios.Cytopathic effect and growth pattern of FAV 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 1075 TCID50/0.1 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 excellent tool for constrution of recombinant fowl adenovirus type Ⅰ .%[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(107.5 TCID50/0.1 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FAV Ⅰ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.