Method for cultivating attenuated strain pilio inactivated vaccine
A poliomyelitis and culture method technology, which is applied to the culture field of poliomyelitis inactivated vaccine, can solve the problem that cell yield and virus yield cannot meet large-scale production, and achieves good safety, immunogenicity and stability. , the effect of reducing the cultivation space
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Embodiment 1
[0024] 1. Tertiary amplification cell culture:
[0025] a. First, the cell seeds are centrifugally cleaned, and inoculated into a 7-liter fermenter at a ratio of minimum requirement culture medium (MEM):Vero cell seeds=500:1, and the culture temperature is controlled at 37°C and pH7.2 , dissolved oxygen 50%, and perfusion began on the third day of culture, and the perfusion flow was finally increased from 1 liter / day to 5 liters / day. After 7 days of culture, the cell proliferation reached 2.13×10 6 cells / ml, end the primary culture, and perform trypsinization;
[0026] b. The cell suspension digested with trypsin in step a is inoculated in a 75-liter fermenter at a ratio of minimum requirement culture medium (MEM): Vero cell seed=500: 1, and the culture temperature is controlled at 37°C and pH7. .2, 50% dissolved oxygen, under the condition of 100 liters of recirculation medium (MEM) volume, begin to carry out recirculation culture on the third day, cultivate 7 days, cell den...
Embodiment 2
[0030] 1. Tertiary amplification cell culture:
[0031] a. First, the cell seeds are centrifugally cleaned, and inoculated into a 10 liter fermenter for cell culture according to the ratio of minimum requirement culture medium (MEM): Vero cell seeds=1000: 1, and the culture temperature is controlled at 35° C. and pH 7.0 , dissolved oxygen 30%, and perfusion began on the third day of culture, and the perfusion flow was finally increased from 1 liter / day to 5 liters / day. After 5 days of culture, the cell proliferation reached 2.93×10 6 cells / ml, end the primary culture, and perform trypsinization;
[0032] b. The cell suspension digested with trypsin in step a is inoculated in an 85-liter fermenter at a ratio of minimum requirement culture medium (MEM): Vero cell seed=1000: 1, and the culture temperature is controlled at 35°C and pH7. .0, dissolved oxygen 40%, under the condition of 100 liters of recirculation medium (MEM) volume, begin to carry out recirculation culture on the...
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