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

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.