应用正交试验优化猪细小病毒N株在IBRS-2细胞转瓶培养

Optimization of IBRS-2 Cells Culturing Condition for Cultivation of Porcine Parvovirus N Strain Using Roller Bottle

利用正交试验对猪细小病毒(PPV)N株在IBRS-2细胞转瓶培养中的培养条件进行优化,为PPV N株弱毒疫苗的大量培养工艺提供参考。设细胞培养液pH值(7.0、7.2和7.4)、接毒时间(0、24和48 h)、接毒剂量(0.10%、1.00%和10.00%)、收毒时间(48、72和96 h)等因子和水平。结果表明,PPV N株转瓶培养的最佳培养条件组合为细胞培养液pH值7.2,采用同步接毒,接毒剂量为0.10%,收毒时间为接毒后96 h。其中收毒时间是影响PPV N株毒价的最主要因素。

The orthogonal design was used to optimize the culture conditions, of PPV N strain in IBRS-2 cell cultivated in roller bottles to pro- vide reference for the mass culture of PPV N attenuated vaccine. Affected factors were as follows ： the pH value （7.0, 7.2, 7.4）, the time of inoculation （0, 24, 48h）, the dose of toxin （0.10 % , 1.00 %, 10.00 % ） and the time of harvesting （48, 72, 96 h）. Results showed that the best culture conditions of PPV N strain IBRS-2 cells cultivated in roller bottles were as follows： pH =7.2, synchronization inocula- tion, 0.10 % inoculation dose, harvesting the virus solution 96 h after inoculation. Virus solution harvest time is the most important factor affecting the virus titer of PPV N strain.