Method for production of porcine epidemic diarrhea virus
A technology for porcine epidemic diarrhea and a production method, which is applied to the production technology field of the rolling bottle culture method, can solve the problems of being easily polluted by the environment, pollute the quality of vaccines, and high production costs, and achieve the advantages of reducing production costs, uniform product quality, and short production cycles. Effect
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[0024] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.
[0025] (1) Choose a bioreactor as the means of cultivation: 75L bioreactor.
[0026] (2) Microcarriers are selected as the carrier for cell-attached growth: Microcarrier Cytodex-2
[0027] (3) Cleaning and sterilization methods of microcarriers: 1) Weigh 10 g / L of Cytodex-2 microcarriers and soak in 2L of PBS solution for three hours. 2) Wash 3 times with 2L PBS each time. 3) Add 2L of PBS solution to soak the microcarriers, and steam sterilize at 121° C. for 30 minutes.
[0028] (4) Select VERO cells as cells for making seedlings:
[0029] (5) Passage and culture of cells for seedling production: the above cells were digested and passaged by EDTA-trypsin cell dispersion (Hank's solution containing 0.25% trypsin (specification: 1:250), 0.02% EDTA). 92% DMEM solution, 8% calf serum, 200 IU / ml sodium penicillin and streptomycin sulfate, and t...
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