Method for producing ring II type DBN-SX07 strain virus for pigs by seroculturing PK-15 cells
A technology of DBN-SX07 and PK-15, which is applied in the field of veterinary biology, can solve problems such as unexamined, and achieve the effects of reducing production costs, expanding production scale, and improving quality
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Embodiment 1
[0036] Embodiment 1: the method for cultivating PK-15 cells with low serum to produce porcine circular type II type DBN-SX07 strain virus, it may further comprise the steps:
[0037] S1. Subculture and culture of cells for preparation:
[0038] Take a T75 flask and culture a monolayer of PK-15 cells, digest the cells with EDTA-trypsin cell dispersion solution, the EDTA-trypsin cell dispersion solution is a mixture of 0.15% trypsin and 0.02% EDTA in Hank's solution solution; use cell growth solution to blow and disperse cells, add 20ml cell growth solution, and place PK-15 cells in a carbon dioxide incubator at a temperature of 37±2°C for 72 hours. When a good cell monolayer is formed, carry out amplified culture. The above-mentioned cell growth medium is DMEM culture medium with 10% serum content;
[0039] S2. Adapting cells to low serum medium: including the following sub-steps:
[0040] S21. Domestication of first generation cells:
[0041] Inoculate the PK-15 cells expan...
Embodiment 2
[0055] Embodiment 2: the method for culturing PK-15 cells with low serum to produce porcine circotype II type DBN-SX07 strain virus, it may further comprise the steps:
[0056] S1. Subculture and culture of cells for preparation:
[0057] Take a T75 flask and culture a monolayer of PK-15 cells, digest the cells with EDTA-trypsin cell dispersion solution, the EDTA-trypsin cell dispersion solution is a mixture of 0.15% trypsin and 0.02% EDTA in Hank's solution Blow and disperse cells with cell growth medium, add 20ml of cell growth medium, place PK-15 cells in a carbon dioxide incubator at a temperature of 37±2°C for 72 hours, and carry out amplified culture when a good cell monolayer is formed. The above cell growth medium is 10% serum content MEM culture medium;
[0058] S2. Adapting cells to low serum medium: including the following sub-steps:
[0059] S21. Domestication of first generation cells:
[0060] The Marc-145 cells expanded and cultivated in step S1 were inoculat...
Embodiment 3
[0074] Embodiment 3: low serum medium acclimates PK-15 cell effect
[0075] Use T75 cell flasks, 37 ℃ static culture adapted to the fourth generation of PK-15 cells in low serum medium, the ratio of serum addition is 1%, 2%, 3%, 4%, 5% five gradients, and the serum content used is set to The PK-15 cells cultured in 10% DMEM was used as the control group, and the results were as follows: figure 1 , figure 2 , image 3 , Figure 4 , Figure 5 , Figure 6 See Table 1 for the indicators during the cell culture process.
[0076] Table 1: Effects of MD series low-serum medium on acclimatization of PK-15 cells
[0077]
[0078] It can be seen from Table 1 that the average cell yield, cell activity and average specific growth rate of the experimental group were higher than those of the control group.
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