A kind of cultivation method of porcine epidemic diarrhea virus
A technology of porcine epidemic diarrhea and culture methods, applied in the direction of microorganism-based methods, viruses/bacteriophages, biochemical equipment and methods, etc., which can solve the problems of difficult cultivation of wild strains
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Embodiment 1
[0044] (1) Preparation of DMEM medium: Dissolve DMEM dry powder in deionized water, stir until completely dissolved, adjust the pH value to 7.2, and use a sterile filter membrane of ≤0.22 μm for sterile filtration, and place it in a room of ≤4°C Refrigerator ready for use;
[0045] (2) adding fetal calf serum accounting for 1-8% of the volume of the DMEM medium in the above-mentioned prepared DMEM medium, and placing the DMEM medium in a culture vessel for culturing Vero monolayer cells;
[0046] (3) Discard the DMEM medium in the culture container with the monolayer cells formed, and wash with washing liquid for 2-3 times; the washing liquid is sterile deionized water.
[0047] (4) Discard washing liquid, add porcine trypsin, cell affinity agent and the DMEM substratum in step (1) in the culture vessel that forms monolayer cell, wherein the consumption of porcine trypsin is DMEM substratum volume consumption 0.3% of DMEM medium volume, and the amount of cell affinity agent is ...
Embodiment 2
[0050] (1) Preparation of DMEM medium: Dissolve DMEM dry powder in deionized water, stir until it is completely dissolved, adjust the pH value to 6.8, and perform sterile filtration with a sterile filter membrane of ≤0.22 μm, and place it in a room of ≤4°C Refrigerator ready for use;
[0051] (2) adding fetal calf serum accounting for 1-8% of the volume of the DMEM medium in the above-mentioned prepared DMEM medium, and placing the DMEM medium in a culture vessel for culturing Vero monolayer cells;
[0052] (3) Discard the DMEM medium in the culture container with the monolayer cells formed, and wash with washing liquid for 2-3 times; the washing liquid is sterile deionized water.
[0053] (4) Discard washing liquid, add porcine trypsin, cell affinity agent and the DMEM substratum in step (1) in the culture vessel that forms monolayer cell, wherein the consumption of porcine trypsin is DMEM substratum volume consumption 0.1% of DMEM medium volume, and the amount of cell affin...
Embodiment 3
[0056] (1) Preparation of DMEM medium: Dissolve DMEM dry powder in deionized water, stir until it is completely dissolved, adjust the pH value to 7.8, and use a sterile filter membrane of ≤0.22 μm for sterile filtration, and place it in a room of ≤4°C Refrigerator ready for use;
[0057] (2) adding fetal calf serum accounting for 1-8% of the volume of the DMEM medium in the above-mentioned prepared DMEM medium, and placing the DMEM medium in a culture vessel for culturing Vero monolayer cells;
[0058] (3) Discard the DMEM medium in the culture container with the monolayer cells formed, and wash with washing liquid for 2-3 times; the washing liquid is sterile deionized water.
[0059] (4) Discard washing liquid, add porcine trypsin, cell affinity agent and the DMEM substratum in step (1) in the culture vessel that forms monolayer cell, wherein the consumption of porcine trypsin is DMEM substratum volume consumption 0.5% of DMEM medium volume, and the amount of cell affinity a...
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Abstract
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