Method for establishing influenza virus infected cell models by using primary cells of tupaia belangeri
A technology for influenza virus infection, primary cells, applied in the field of animal cell lines, to achieve the effect of uniform expression
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Embodiment 1
[0026] Example 1: Using tree shrew primary cells to establish influenza virus infection cell model
[0027] (1) Cultivation of tree shrew primary cells:
[0028] a. Cultivation of primary cells of tree shrew trachea:
[0029] The trachea of the tree shrew was exposed, the lungs and the trachea were tied tightly with surgical rope, and a collagenase digestion solution containing 400 U / ml penicillin and 0.1% mass concentration was injected into the trachea at the mouth end. Tighten the trachea at the end of the mouth after injection, fill the trachea with collagenase digestion solution, cut off the ends other than the binding place, and take out the trachea. Put it in a petri dish containing 400U / ml penicillin PBS, and digest it for 24 hours at a temperature of 37°C. After enzymatic digestion, cut off one end of the sealed trachea, and flow out the collagenase digestion solution and the primary cells of the tree shrew trachea. Centrifuge them at 3000 rpm for 8-12 minutes to remove ...
Embodiment 2
[0036] Example 2: Cell virus sensitivity test
[0037] In the tree shrew tracheal primary cells, tree shrew lung primary cells and tree shrew renal medulla primary cells for virus culture, three tree shrews were taken out at three different time points at 24 hours, 48 hours, and 72 hours. The supernatant of the next generation of cells was tested for virus titer to determine the virus sensitivity of the cells to influenza virus A / PR / 8 / 34 (H1N1). The experimental results are shown in Table 1.
[0038] Table 1 Virus titers in the supernatant of primary tree shrew cells at different time points
[0039]
[0040]
[0041] It can be seen from Table 1 that after 72 hours, the viruses amplified by the primary cells of the three tree shrews were all greater than 2LgTCID 50 / 0.1mL (i.e. 100TCID 50 / 0.1mL), indicating that the primary cells of the three tree shrews are sensitive to influenza virus A / PR / 8 / 34 (H1N1) and have the effect of amplifying influenza virus. Using tree shrew primary ...
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