无血清悬浮MDCK细胞库的建立及检定

Establishment and control of serum-free suspended MDCK cell bank

目的建立无血清悬浮MDCK细胞的主代细胞库及工作代细胞库,并进行检定。方法将购自欧洲细胞株/微生物保藏中心(European Collection of Authenticated Cell Cultures,ECACC)的无血清悬浮MDCK细胞扩大培养并建立主代细胞库和工作代细胞库,按照《中国药典》三部(2015版)通则、生物制品生产检定用动物细胞基质制备及检定规程中的规定,对建立的细胞库进行检定,并进行流感病毒敏感性检测。结果建立的无血清悬浮MDCK细胞的主代细胞库及工作代细胞库细胞形态与文献中记录的悬浮MDCK细胞形态一致,各项检定结果均符合《中国药典》三部(2015版)中的规定,且对流感病毒保持了高敏感性。结论成功建立了无血清悬浮MDCK细胞库,为细胞基质流感疫苗的研究奠定了基础。

Objective To establish and control a master cell bank(MCB)and a work cell bank(WCB)of serum-free suspended MDCK cells.Methods Serum-free suspended MDCK cells purchased from the European Collection of Authenticated Cell Cultures(ECACC)were subjected to expanded culture,based on which MCB and WCB were established and subjected to control tests according to the General Requirements and the Requirements for Animal Matrix Cells for Production and Control of Biologics in Chinese Pharmacopoeia(Volume Ⅲ,2015 edition),and tested for sensitivity to influenza virus.Results The cell morphologies in established MCB and WCB of serum-free suspension MDCK cells were consistent with those of suspended MDCK cells published in literatures,while the control test results met the requirements in Chinese Pharmacopoeia(Volume Ⅲ,2015 edition).Meanwhile,the cells from MCB and WCB were highly sensitive to influenza virus.Conclusion The serum-free suspended MDCK cell bank was established,which laid a foundation of study on cell-based influenza virus vaccine.