Method for producing porcine reproductive and respiratory syndrome JXA1-R strain virus by using low serum cultured Marc-145 cells
A JXA1-R, low-serum technology, applied in the field of veterinary biology, can solve problems such as no, and achieve the effect of increasing production costs
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Embodiment 1
[0033] Embodiment 1: the method that low serum culture Marc-145 cell produces porcine reproductive and respiratory syndrome JXA1-R strain virus, it comprises the following steps:
[0034] S1. Subculture and culture of cells for preparation:
[0035] Take a T75 flask and culture a monolayer of Marc-145 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; blow and disperse the cells with cell growth solution, add 20ml of cell growth solution, and place Marc-145 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;
[0036] S2. Adapting cells to low serum medium: including the following sub-steps:
[0037] S21. Domestication of first generation cells:
[0038]Inoculate the Ma...
Embodiment 2
[0049] Embodiment 2: the method that low serum culture Marc-145 cell produces porcine reproductive and respiratory syndrome JXA1-R strain virus, it comprises the following steps:
[0050] S1. Subculture and culture of cells for preparation:
[0051] Take a T75 flask and culture a monolayer of Marc-145 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 the cells with cell growth medium, add 20ml of cell growth medium, place Marc-145 cells in a carbon dioxide incubator at a temperature of 37±2°C for 72 hours, and perform amplified culture when a good cell monolayer is formed. The above-mentioned cell growth medium is MEM culture medium with 10% serum content;
[0052] S2. Adapting cells to low serum medium: including the following sub-steps:
[0053] S21. Domestication of first generation cells:
[0054] The Marc-145 cells expanded and c...
Embodiment 3
[0065] Example 3: Effect of low serum medium on acclimating Marc-145 cells
[0066] Use T75 cell flasks, 37 ℃ static culture adapted to the fourth generation of Marc-145 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 Marc-145 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.
[0067] Table 1: Effects of MD series low-serum medium on acclimatization of Marc-145 cells
[0068]
[0069] 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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