无血清微载体培养MDCK细胞和甲型流感病毒H1N1的条件优化

 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.

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

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 in...