Preparation method of animal interferon
A technology of interferon and animals, which is applied in the field of animal medicine, can solve the problems of low interferon content and no reports, and achieve the effects of low dosage, low preparation cost and simple production process
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Embodiment 1
[0020] The concrete steps of a kind of preparation method of animal interferon are as follows:
[0021] (1) Chicken spleen lymphocyte preparation
[0022] Aseptically take the spleen of the experimental chicken, prepare the lymphocyte suspension with the spleen, add 10 μg / ml kanamycin, dilute the lymphocyte suspension with the prepared culture medium, and dilute the chicken spleen lymphocyte concentration to 1×106 per ml . The isolation of chicken spleen lymphocytes should be carried out within 48 hours after collection. The number of living cells should reach more than 90%. And the lymphocyte monolayer was subcultured; the chicken spleen lymphocyte concentration was diluted to contain 1×10 per ml 6 . Isolation of chicken spleen lymphocytes should be performed within 48 hours of preparation. The number of living cells should reach more than 90%.
[0023] (2) induced virus
[0024] Sendai virus is used, and the hemagglutination titer can only be put into production when ...
Embodiment 2
[0044] The concrete steps of a kind of preparation method of animal interferon are as follows:
[0045] (1) Aseptically take experimental chicken spleen, prepare lymphocyte suspension with spleen, 10 μg / ml penicillin and streptomycin, dilute cell sediment with prepared 1640 medium, so that chicken spleen lymphocyte concentration is diluted to contain 1×10 7 .
[0046] (2) Add 8% chicken plasma to chicken spleen lymphocyte suspension and mix well.
[0047] (3) A small amount of interferon was added to the above chicken spleen lymphocyte suspension to make the final concentration 200-300 IU / ml, and cultured in a water bath at 37° C. for 1 hour.
[0048] (4) Add astragalus polysaccharides to make the final concentration 25 μg / ml; incubate in a water bath at 37° C. for 2 hours.
[0049] (5) Add 512 hemagglutination units / ml of Sendai virus to the activated chicken spleen lymphocyte suspension for induction, and culture on a shaker at 37° C. for 12-24 hours.
[0050] (6) When t...
Embodiment 3
[0055] A preparation method of animal interferon, the specific steps are as follows:
[0056] (1) Aseptically collect experimental chicken spleen, prepare lymphocyte suspension with spleen, add 10 μg / ml kanamycin, and place in refrigerator at 4.0°C. Dilute the cell pellet with the prepared culture medium, so that the chicken spleen lymphocyte concentration is diluted to 1×10 per ml 5 .
[0057] (2) A small amount of interferon was added to the chicken spleen lymphocyte suspension to make the final concentration 800 IU / ml, and cultured in a water bath at 37°C for 2 hours.
[0058] (3) Astragalus polysaccharide was added to make the final concentration 50 μg / ml, and cultured in a water bath at 37° C. for 2 hours.
[0059] (4) Add 256-1280 hemagglutination units / ml Sendai virus again in the activated chicken spleen lymphocyte suspension to induce, and add the DMEM medium containing 5% bovine serum in the chicken spleen lymphocyte suspension simultaneously, 37 Cultivate on a shak...
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