Cloning and analyzing of MDCK cells for H5N1 influenza vaccine production
10.3760/cma.j.issn.0254-5101.2019.02.001
- VernacularTitle:适用于生产H5N1流感疫苗的MDCK亚克隆细胞的筛选及检定
- Author:
Xiaoyuan HUANG
1
,
2
;
Jiayou ZHANG
;
Yang LIU
;
Wei ZHAO
;
Jinrong SHI
;
Xinru LE
;
Gang YU
;
Xixin HAN
;
Xiaoming YANG
Author Information
1. 武汉生物制品研究所有限责任公司流感疫苗课题组 430207
2. 国家联合疫苗工程技术研究中心
- Keywords:
MDCK cell;
H5N1 influenza virus;
Tumorgenicity and oncogenicity
- From:Chinese Journal of Microbiology and Immunology
2019;39(2):81-87
- CountryChina
- Language:Chinese
-
Abstract:
Objective To screen a Madin-Darby canine kidney (MDCK) cell line for H5N1 influ-enza virus isolation and to evaluate its safety in vaccine production. Methods MDCK cells were cloned by the method of limiting dilution. Hemagglutination test was used to screen MDCK cells that were suitable for H5N1 influenza virus production. Tests for analyzing the characteristics, extraneous agents, endogenous agents and tumorigenicity of MDCK cells were performed according to Chinese Pharmacopeia Volume Ⅲ. Results A total of 108 MDCK cell lines were obtained and three of them were selected after hemagglutina-tion test. G1 cells were chosen following further screening with tumorigenicity test and receptor abundance analysis. The average number of chromosomes of the MDCK-G1 cells was 78±4. No bacteria, fungi or myco-plasma contamination was detected. In experimental group, each nude mouse was injected with 1×107/ml viable cells to observe their tumorigenicity. Twelve weeks after cell injection, no node was found at injection sites or in gross anatomy. There was no significant difference between the experimental and negative control groups. The result of the tumorigenicity test was negative. No node formation was found after injecting nude mice with cell lysate or cellular DNA collected from equivalent amount of cells. It was indicated that the MDCK-G1 cells were of low-oncogenic potential. Conclusions The MDCK-G1 cell line could be used as a substrate to produce H5N1 influenza virus vaccine.
