张严予, 马磊, 沈霏, 周文晶, 周玉波, 王微, 姜述德, 李卫东, 廖国阳. 无血清微载体培养MDCK细胞和甲型流感病毒H1N1的条件优化[J]. 云南大学学报(自然科学版), 2011, 33(3): 340-344 .
引用本文: 张严予, 马磊, 沈霏, 周文晶, 周玉波, 王微, 姜述德, 李卫东, 廖国阳. 无血清微载体培养MDCK细胞和甲型流感病毒H1N1的条件优化[J]. 云南大学学报(自然科学版), 2011, 33(3): 340-344 .
ZHANG Yan-yu, MA Lei, SHEN Fei, ZHOU Wen-jing, ZHOU Yu-bo, WANG Wei, JIANG Shu-de, LI Wei-dong, LIAO Guo-yang. Optimizing the serum-free micro-carrier culture condition of MDCK cells and influenza Virus H1N1[J]. Journal of Yunnan University: Natural Sciences Edition, 2011, 33(3): 340-344 .
Citation: ZHANG Yan-yu, MA Lei, SHEN Fei, ZHOU Wen-jing, ZHOU Yu-bo, WANG Wei, JIANG Shu-de, LI Wei-dong, LIAO Guo-yang. Optimizing the serum-free micro-carrier culture condition of MDCK cells and influenza Virus H1N1[J]. Journal of Yunnan University: Natural Sciences Edition, 2011, 33(3): 340-344 .

无血清微载体培养MDCK细胞和甲型流感病毒H1N1的条件优化

Optimizing the serum-free micro-carrier culture condition of MDCK cells and influenza Virus H1N1

  • 摘要: 血清微载体培养MDCK细胞,并接种流感病毒H1N1,优化培养条件,为细胞流感疫苗的工艺研发奠定基础.采用不同的细胞接种量在50mL无血清微载体搅拌瓶中培养MDCK细胞,并接种甲型流感病毒H1N1,检测不同pH值和TPCK-胰酶含量的病毒培养液,不同病毒接种量,补加TPCK-胰酶,以及不同收毒时间对血凝效价的影响.以1.0×105mL-1的MDCK细胞数量接种到无血清微载体上,病毒培养液pH值为7.2~7.4范围内,TPCK-胰酶质量浓度为1.0μg/mL,接种后不补加胰酶,病毒接种量MOI=1.0,并在72h收获,最高血凝效价达到512.由此获得了在无血清为载体上培养MDCK细胞和甲型流感病毒H1N1的适宜条件.

     

    Abstract: The article is about the optimum culture conditions for micro-carrier cultivating MDCK cells and influenza virus H1N1 proliferation in Serum-Free Medium,which is the basement for the cell influenza vaccine in the future.We cultivated MDCK cell in VP-SFM,detected the effect of MDCK cell growth with different concentration of cell inoculation,then cultivated influenza virus H1N1 in MDCK cell and also detected the effect of different pH value,TPCK-Trypsin content,MOI,whether or not adding TPCK-Trypsin after inoculating influenza virus and different virus harvest time.When the cell quantity is 1.0×105mL-1,MDCK cell can grow for 5—6 days,which cell status are in good condition.After inoculating influenza virus H1N1 to MDCK cells,when pH value is between 7.2—7.4,TPCK-Trypsin content is 1.0μg/mL,MOI is 1.0, and virus harvest time is 72h,the hemagglutination titer is up to 512.In the selected suitable conditions,we could use microcarrier culture method for cultivation of MDCK cells and influenza virus H1N1 in Serum-Free Medium on a large scale in the future.

     

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