基于悬浮型MDCK细胞冻存和快速复苏的一种流感病毒培养方法

A method for cultivating and isolating influenza virus based on cryopreservation and rapid revival of MDCK cells cultured in suspension

本文使用一种基于高分子聚合物的细胞保护剂(CCM046)冻存和复苏悬浮型MDCK细胞,并探索开发一种更加便捷的流感病毒培养方法。结果显示,CCM046细胞冻存液可稳定的于-80℃条件下保存悬浮型MDCK细胞,且细胞活力稳定,可在复苏后快速恢复细胞增殖。在细胞复苏24 h后进行攻毒实验,培养48 h后所产生的流感病毒含量基本与持续培养的新鲜细胞相同,初步证明该方法可用于建立一种快速、便捷的使用MDCK细胞培养和分离流感病毒的技术方法。

MDCK cells cultured in suspension were subjected to cryopreservation and subsequent revival using a cryopreservation medium supplemented with high molecular weight polymers.This approach was employed to assess the feasibility of developing a streamlined procedure for cultivating influenza virus.The findings indicated that the cryopreservation solution effectively maintained the viability of MDCK cells cultured in suspension at-80 degrees Celsius,ensuring consistent cell viability and enabling rapid cellular proliferation upon revival.Furthermore,following a 48-hour influenza virus infection period,the viral yield obtained from cells recovered 24 hours after cryopreservation was comparable to that from continuously cultured fresh cells.These preliminary findings imply that this method holds promise for the establishment of a rapid and expedient protocol for both influenza virus cultivation and isolation,built upon MDCK cell cultures.