Stability of Suspended MDCK Cells for Avian Influenza Virus Production

doi:10.13560/j.cnki.biotech.bull.1985.2020-0434

Abstract

Abstract:

The aim of this study is to determine the stability of growing and virus producing for suspension MDCK cells during long-term passaging culture. Suspended MDCK cells were subcultured for more than 200 d. Taking suspended MDCK cells at different passaging time points,their growth patterns and the difference in virus production of H9N2 avian influenza virus（H9N2-AIV）were investigated. The effects of different process parameters and conditions such as medium dilution ratios and MOI on the production of H9N2-AIV from the suspended long-term passaging MDCK cells was explored. The reasons for the difference in H9N2-AIV production capacity between the lowest passaged cells（P1）and the highest passaged cells（P111）were preliminarily investigated from two aspects,i.e. the intracellular ROS level and cell cycle. The results showed that the MDCK cells at the different passages presented similar morphology,and insignificant difference was observed concerning cell growth between all subcultures and batch cultures,which indicated that the growth of the MDCK cells were stable during long-term passaging. The HA titer increased with elevated cell passaging time,the HA titer of P111 was about 0.5 Log₂HAU/100 μL higher than that of P1. Meanwhile,P111 cells produced higher virus than P1 cells under different tested processes,indicating that the H9N2-AIV production capacity of the suspended MDCK cells had slightly increased with the increase of cell passaging time. It was also found that P111 cells had lower ROS level and higher proportion of G0/G1-phase cells,which may be the main reasons for the increased yield of H9N2-AIV from MDCK cells.

Key words: suspended MDCK cells, avian influenza virus, cell line stability, ROS, cell cycle

BAI Chun-li, YE Qian, JI A-mei, LIU Xu-ping, ZHANG Xu, LIU Zhi-liang, ZHU Ming-long, ZHAO Liang, TAN Wen-song. Stability of Suspended MDCK Cells for Avian Influenza Virus Production[J]. Biotechnology Bulletin, 2020, 36(10): 72-79.

Figures/Tables 8

References 21

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Passage	细胞周期分布/%
	G0/G1	S	G2/M
P1	46.16±0.67	24.18±0.43	26.44±1.31
P111	57.58±0.36	19.36±0.29	20.58±0.33

Passage	细胞周期分布/%
	G0/G1	S	G2/M
P1	46.16±0.67	24.18±0.43	26.44±1.31
P111	57.58±0.36	19.36±0.29	20.58±0.33