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Method for culturing goose parvovirus by using goose embryo fibroblast line

A fibroblast and gosling plague virus technology, applied to embryonic cells, microbial-based methods, animal cells, etc., can solve the problems of low production efficiency, achieve stable immunogenicity, improve virus culture efficiency, and flexibly arrange Effect

Inactive Publication Date: 2015-03-25
黑龙江省兽医科学研究所
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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for cultivating gosling plague virus with goose embryo fibroblast cell line in order to solve the problem of low production efficiency of the existing gosling plague virus cultivation method

Method used

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Embodiment Construction

[0013] The specific steps of the method for cultivating gosling plague virus with goose embryo fibroblast cell line are as follows:

[0014] (1) Cultivation of goose embryo fibroblast primary cells: take 12-day-old well-developed goose embryos, use iodine wine cotton and alcohol cotton to sterilize the air chamber of the eggshell, take out the goose embryos aseptically, remove the head, limbs and internal organs, Put it into a sterilized glass dish, wash the embryo body three times with DMEM solution, cut it into small tissue pieces the size of rice grains with sterilized scissors, wash it twice with DMEM solution, and then add 0.25% trypsin solution (per goose Embryo plus 3~5ml), digest in 37.5℃ water bath for 5~10min, suck out the trypsin solution, wash twice with DMEM solution, then add DMEM solution containing 8% calf serum, pH value 7.0~7.5 to blow and disperse the digested cells After filtration, a cell suspension containing 1 to 1.5 million viable cells per milliliter ...

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Abstract

The invention provides a method for culturing goose parvovirus by using a goose embryo fibroblast line, belongs to the field of veterinary biological products, and solves the problem of low production efficiency of an existing goose parvovirus culturing method. The method comprises the following steps: culturing primary cells, namely cutting a goose embryo which well develops into small pieces, washing, digesting, blowing and dispersing digestive cells to obtain a cell suspension, culturing until the cells form complete monolayers so as to obtain the primary cells; establishing a goose embryo fibroblast line, namely removing a nutrient solution from the primary cells of the goose embryo fibroblast, washing, digesting and culturing until complete monolayer cells are formed to obtain F1-generation cells which can be passed by three generations; reproducing and harvesting, namely inoculating F1-F4-generation cells which form 70 percent of monolayer, and harvesting reproduced viruses when over 75 percent of cells is diseased. According to the method, the amount of harvested virus liquid is 255 times of the dose of primary cell inoculation culturing, and the virus titer of the virus liquid is 10-100 times higher than that of a virus liquid which is inoculated, cultured and harvested when 100 percent of monolayer is formed.

Description

technical field [0001] The invention belongs to the field of veterinary biological products, and in particular relates to a method for cultivating gosling plague virus. Background technique [0002] my country's goose farming industry has developed rapidly in recent years, and the amount of breeding has increased year by year, which has played a positive role in increasing farmers' income and adjusting the industrial structure. Goose products are considered pure green food, and the economic value of its by-products is also high. The market gap is large and the economic benefits are obvious. These conditions have further promoted the rapid development of the goose industry. However, to ensure the stable and healthy development of the goose industry, a set of scientific and effective disease prevention and control measures must be established. [0003] Gosling plague (GP) is an important infectious disease that endangers the goose industry at present. The disease has caused s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N5/073C12R1/93
Inventor 刘力威李洪彬黄宇翔张军王志强邹跃杨旭东陈亮
Owner 黑龙江省兽医科学研究所
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