Method for proliferating avian influenza virus by adopting MDCK cell line

• Main Authors: ZHAN XUANZI, SHI WEISONG, ZHENG CAILI, LU SHAOHUA, KONG FENGYING, XIE YUHONG, LIU DECHENG
• Format: Patent
• Language: Chinese; English
• Published: 15.09.2020
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; HUMAN NECESSITIES; HYGIENE; MEDICAL OR VETERINARY SCIENCE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS ORMEDICINAL PREPARATIONS; SPIRITS; VINEGAR; WINE

The invention relates to a method for proliferating avian influenza virus by adopting an MDCK cell line. The method comprises the following steps: domesticating adherent MDCK cells into a serum-free full-suspension culture type MDCK cell line which is stably proliferated, inoculating a bioreactor with the MDCK cell line, completing primary cell culture, preparing avian influenza H5N1 subtype virusseed viruses for suspension MDCK cell culture, inoculating suspension MDCK cells with the avian influenza H5N1 subtype virus seed viruses, performing culturing, and separating and purifying virus particles. The cells obtained by domestication are stable in passage, good in cell morphology, high in quality, capable of keeping sensitivity and biological characteristics to viruses, high in virus multiplication capacity and capable of effectively multiplying avian influenza virus strains. 本发明涉及一种采用MDCK细胞系增殖禽流感病毒的方法，所述方法包括将贴壁的MDCK细胞驯化为稳定增殖的无血清全悬浮培养型MDCK细胞系，然后接种于生物反应器，完成初级细胞培养，制备悬浮MDCK细胞培养用禽流感H5N1亚型病毒种毒，接毒于悬浮MDCK细胞进行培养，分离并