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Preparation method of permanent cell line for multiplying orf virus

A kind of oral ulcer virus and cell line technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, cells modified by introducing foreign genetic materials, etc., can solve time-consuming and financial resources, difficult quality control, and complicated cell preparation procedures And other issues

Active Publication Date: 2014-08-20
NORTHWEST A & F UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] However, there is currently no oral ulcer vaccine on the market for users to use. The reason for this problem is that vaccine manufacturers are unwilling to produce attenuated oral oral vaccine, because the cells needed for the production of this vaccine are calf primary testicular cells, and these cells are prepared The procedures are complicated, the production cost is too high, time-consuming and financial resources are consumed, and it is difficult to achieve effective quality control, and it is difficult for enterprises to obtain relatively rich profits

Method used

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  • Preparation method of permanent cell line for multiplying orf virus
  • Preparation method of permanent cell line for multiplying orf virus
  • Preparation method of permanent cell line for multiplying orf virus

Examples

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Embodiment

[0023] The following examples are used to illustrate the present invention, but do not limit the scope of use of the present invention.

[0024] 1. Isolation and culture of calf testicular cells

[0025] Under sterile conditions, Hank’s solution (double antibody) was used to wash the testis three times, and the epididymis and albuginea were cut off with sterile scissors, leaving the testicular parenchyma. Flush the testicular parenchyma with Hank's solution until there is no blood, and then transfer it to a sterile Erlenmeyer flask.

[0026] Use ophthalmic scissors to cut the testicles into pieces, about 1-3mm in size, rinse with Hank's solution, and let stand for 2-3 minutes. Discard the supernatant, wash repeatedly 3 times, add PH7.6, 0.25% trypsin, the dosage is 2 to 4 times the volume of the tissue, and let it stand overnight at 4°C. The next day, remove the trypsin, add M199 culture medium, blow gently to blow off the cells, centrifuge in a 50mL centrifuge tube, 1000rpm...

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Abstract

The invention relates to a preparation method of a permanent cell line for multiplying an orf virus. The aim that the virus can be stably multiplied in a large amount is achieved by inoculating a calf testicular cell line to the orf virus and a stable cell environment and a standardized culture system for developing and producing an attenuated vaccine of the orf virus are provided. The preparation method comprises the following steps of: firstly, acquiring, separating and culturing a calf testicular cell; secondly, permanently establishing the calf testicular cell; and thirdly, carrying out multiplication culture on the orf virus in the cell line.

Description

1. Technical field: [0001] The invention relates to a method for preparing a permanent cell line for multiplying aphthous ulcer virus, which is a method for preparing a cell line from calf testicular cells, and using the cell line for aphthous aphthous ulcer virus to proliferate. 2. Background technology: [0002] In the background technology, sheep infectious impetigo is commonly called as sheep mouth sore (Orf virus), is a kind of zoonotic infectious disease caused by a mouth sore virus of sheep and goats, and the World Organization for Animal Health (OIE) lists this disease as a need The declared animal disease is listed as a first-class animal disease in my country. The disease is distributed in almost all sheep-raising countries in the world, and mainly affects sheep and goats under natural conditions, and goats are more frequent. The disease is often popular in groups, and lambs aged 3 to 6 months are most susceptible to infection, and adult sheep are less likely to be...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12R1/91
Inventor 陈德坤尚川川李杰罗军
Owner NORTHWEST A & F UNIV
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