Comparison of isolating influenza virus in culture systems of adherent and a suspending cells of MDCK cell line
10.3760/cma.j.cn112866-20230522-00053
- VernacularTitle:悬浮与贴壁MDCK细胞在流感病毒分离培养中的应用评价
- Author:
Xue ZHOU
1
;
Junling YU
;
Sitian YANG
;
Yidan XIA
;
Wanrong LUO
;
Weixi FANG
;
Minhao HU
;
Jun HE
Author Information
1. 安徽省疾病预防控制中心（安徽省公共卫生研究院），合肥 230601
- Keywords:
Influenza virus;
MDCK cell suspension;
Adherent MDCK cell;
Virus isolation
- From:
Chinese Journal of Experimental and Clinical Virology
2023;37(6):637-642
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To compare the differences between suspension and adherent cells of MDCK cell line in the isolation of influenza virus, and to explore the application prospects of MDCK cell suspension.Methods:Determination of viable cell density and cell specific growth rate were recorded by cell count. The WHO recommended vaccine strains were used for virus infection experiments. After five passages, hemagglutination titers were detected, while the sequencing analysis of their HA and NA genes revealed the mutation frequency.Results:The 24-hour and 48-hour viable cell density of the cell suspension was more stable than that of adherent cells. The cell suspension achieved an HA titer of 1∶256 or higher in the third generation, while adherent cells had no titer. In the fourth and fifth generations, one amino acid site mutation was found in the HA gene of H3N2 and BV subtypes of influenza virus cultured in the cell suspension, while no gene mutation was found in adherent cells in two passages. There were no mutations in the whole NA gene.Conclusions:Suspension of MDCK cells have more stable growth and higher efficiency in virus isolation than adherent cells, meanwhile there was a low rate of virus mutation during continuous passage. This study demonstrated the feasibility of this suspension of MDCK cells for influenza vaccine production based on cell culture technology.
