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

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(107.5 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。

Abstract:

[Objective] To explore culture conditions for chicken hepatocellular carcinoma cell line (LMH) and growth pattern of fowl adenovirus typeⅠ-AV208 strain (FAVⅠ-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 FAVⅠ- 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. FAVⅠ- AV208 could be propagated efficiently in LMH cells and high virus titer up to 107.5 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 typeⅠ.