In-vitro preparation method of transfer factors capable of resisting duck viral hepatitis
A technology for duck viral hepatitis and transfer factor, applied in the field of immunology, can solve the problems of inability to play a therapeutic role, limited curative effect, poor pertinence, etc., and achieve the effects of low cost, low production cost and manpower saving.
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Embodiment 1
[0018] A preparation method for in vitro anti-duck viral hepatitis transfer factor, the present embodiment uses chicken peripheral blood as the basic raw material to prepare monolayer lymphocytes, and induces and cultures monolayer lymphocytes through phytohemagglutinin and duck hepatitis virus, thereby promoting To produce a large amount of anti-duck hepatitis virus specific transfer factor in vitro, the specific steps are as follows:
[0019] 1. Collection of anticoagulant blood: Select healthy chickens without infectious diseases, sterilize the blood collection site, use a 50mL sterile syringe to collect anticoagulant blood from the heart, and add 5mL of heparin sodium with a concentration of 1mg / mL;
[0020] 2. Lymphocyte separation: Add anticoagulant blood and lymphocyte separation solution (purchased from Beijing Puli Biological Co., Ltd., and operate according to the instructions) into the sterilized centrifuge tube at a ratio of 1:1, and add the lymphocyte separation so...
Embodiment 2
[0026] A preparation method for in vitro anti-duck viral hepatitis transfer factor, the present embodiment uses chicken peripheral blood as the basic raw material to prepare monolayer lymphocytes, and induces and cultures monolayer lymphocytes through phytohemagglutinin and duck hepatitis virus, thereby promoting To produce a large amount of anti-duck hepatitis virus specific transfer factor in vitro, the specific steps are as follows:
[0027] 1. Collection of anticoagulant blood: Select healthy chickens without infectious diseases, sterilize the blood collection site, use a 50mL sterile syringe to collect anticoagulant blood from the heart, and add 5mL of heparin sodium with a concentration of 1mg / mL;
[0028] 2. Lymphocyte separation: Add anticoagulant blood and lymphocyte separation solution (purchased from Beijing Puli Biological Co., Ltd., and operate according to the instructions) into the sterilized centrifuge tube at a ratio of 1:1, and add the lymphocyte separation so...
Embodiment 3
[0034] An in vitro preparation method of anti-duck viral hepatitis transfer factor, the present embodiment mainly includes the steps of preparing monolayer lymphocytes from chicken spleen, adding phytohemagglutinin and virus induction, and the specific steps are as follows:
[0035] 1. Spleen collection: select healthy chickens without infectious diseases, remove the spleen under aseptic conditions, then wash the blood on the surface and the liquid on the surface of the internal organs with sterilized saline, disinfect the surface with alcohol, and remove the fat tissue and For fascia, use sterile ophthalmic scissors and ophthalmic tweezers to take part of the tissue from the inside of the spleen, put it in a 30mL beaker, then cut it into pieces with a diameter of less than 0.1cm, and wash it three times with pH7.2 Hanks solution Blood and impurities in the tissue, keep the tissue as clean as possible;
[0036] 2. Splenocyte preparation: add 5 times the amount of trypsin s...
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