Method for producing swine fever live vaccine with cell line

Abstract

The invention discloses a method for producing a live swine fever vaccine by using a cell line. The present invention comprises the following technical steps: (1) selecting a cell line as the cells for making seedlings; (2) subculture and cultivation of cells for making seedlings; (3) breeding of cytotoxic species; (4) breeding of venom for making seedlings; 5) Mixing seedlings, subpackaging and freeze-drying. The invention has the advantages of simple and stable production process, easy operation, high virus content, small difference between batches, easy quality control, and can significantly improve the yield and quality of vaccines. The live swine fever vaccine produced by the invention has good safety and high immune efficacy, and has complete immune protection against the virulent attack of swine fever.

Description

technical field

[0001] The invention belongs to the technical field of veterinary biological products, and more specifically relates to a method for producing live swine fever vaccine with cell lines. Background technique

[0002] The cells used in the production of attenuated CSF vaccine in my country are primary bovine testicular cells. Since BVD / MDV disease is relatively common in cattle herds in my country, the use of primary cells of bovine testis to produce vaccines may easily cause cell exogenous virus contamination, and the titer of toxin production is not high, and there are large batch-to-batch differences, which will bring great benefits to the improvement of vaccine production and efficacy. difficulty. Contents of the invention

[0003] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a method for producing live vaccines against swine fever with cell lines. The metho...

Examples

Embodiment 1

[0027] (1) Selection of cells for seedling preparation: select porcine testis (ST) cell line for use;

[0028] (2) Passage and culture of cells for seedling production: ST cell line is digested and passed by EDTA-trypsin cell dispersion liquid, and then cultured with cell growth medium at 36°C. When a good monolayer is formed, it is used for continued passage or inoculation Virus;

[0029] (3) Propagation of cytotoxic species: use cell maintenance solution to make 0.3% virus suspension from fresh spleen venom, inoculate a single layer of well-grown ST cell line, and continue culturing at 36°C. Harvest and change the medium every 4 days, take the second harvest and third harvest of cell culture venom as the poisonous species for production;

[0030] Identification of cytotoxic species: The identification of cytotoxic species fully complies with the standard of attenuated strains of classical swine fever, which is safe for pigs and has no side effects. Each 1ml of cytotoxic spe...

Embodiment 2

[0038] (1) Selection of cells for seedling preparation: select porcine kidney (PK15) cell line for use;

[0039] (2) Passage and culture of cells for seedling production: PK15 cell line was digested and passed by EDTA-trypsin cell dispersion liquid, and continued to culture at 37°C with cell growth liquid. When a good monolayer was formed, it was used for continued passage or virus inoculation;

[0040] (3) Propagation of cytotoxic species: use cell maintenance solution to make 0.4% virus suspension from fresh spleen poison, inoculate a single layer of well-growing PK15 cell line, and place at 37° C. to continue culturing. Harvest and change the medium every 5 days, take the second harvest and third harvest of cell culture venom as the poisonous species for production;

[0041] Identification of cytotoxic species: The identification of cytotoxic species fully complies with the standard of attenuated strains of classical swine fever, which is safe for pigs and has no side effec...

Embodiment 3

[0049] (1) Selection of cells for seedling preparation: select porcine kidney (IBRS-2) cell line for use;

[0050] (2) Passage and culture of cells for seedling production: IBRS-2 cell line was digested and passed by EDTA-trypsin cell dispersion solution, and continued to culture at 37°C with cell growth medium. When a good monolayer was formed, it was used for continued passage or inoculation Virus;

[0051] (3) Propagation of cytotoxic species: use cell maintenance solution to make 0.5% virus suspension from fresh spleen venom, inoculate a single layer of well-growing IBRS-2 cell line, and place it at 37°C to continue culturing. Harvest and change the medium every 4 days, take the second harvest and third harvest of cell culture venom as the poisonous species for production;

[0052] Identification of cytotoxic species: The identification of cytotoxic species fully complies with the standard of attenuated strains of classical swine fever, which is safe for pigs and has no s...

technical field

[0001] The invention belongs to the technical field of veterinary biological products, and more specifically relates to a method for producing live swine fever vaccine with cell lines. Background technique

[0002] The cells used in the production of attenuated CSF vaccine in my country are primary bovine testicular cells. Since BVD / MDV disease is relatively common in cattle herds in my country, the use of primary cells of bovine testis to produce vaccines may easily cause cell exogenous virus contamination, and the titer of toxin production is not high, and there are large batch-to-batch differences, which will bring great benefits to the improvement of vaccine production and efficacy. difficulty. Contents of the invention

[0003] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a method for producing live vaccines against swine fever with cell lines. The metho...

Examples

Embodiment 1

[0027] (1) Selection of cells for seedling preparation: select porcine testis (ST) cell line for use;

[0028] (2) Passage and culture of cells for seedling production: ST cell line is digested and passed by EDTA-trypsin cell dispersion liquid, and then cultured with cell growth medium at 36°C. When a good monolayer is formed, it is used for continued passage or inoculation Virus;

[0029] (3) Propagation of cytotoxic species: use cell maintenance solution to make 0.3% virus suspension from fresh spleen venom, inoculate a single layer of well-grown ST cell line, and continue culturing at 36°C. Harvest and change the medium every 4 days, take the second harvest and third harvest of cell culture venom as the poisonous species for production;

[0030] Identification of cytotoxic species: The identification of cytotoxic species fully complies with the standard of attenuated strains of classical swine fever, which is safe for pigs and has no side effects. Each 1ml of cytotoxic spe...

Embodiment 2

[0038] (1) Selection of cells for seedling preparation: select porcine kidney (PK15) cell line for use;

[0039] (2) Passage and culture of cells for seedling production: PK15 cell line was digested and passed by EDTA-trypsin cell dispersion liquid, and continued to culture at 37°C with cell growth liquid. When a good monolayer was formed, it was used for continued passage or virus inoculation;

[0040] (3) Propagation of cytotoxic species: use cell maintenance solution to make 0.4% virus suspension from fresh spleen poison, inoculate a single layer of well-growing PK15 cell line, and place at 37° C. to continue culturing. Harvest and change the medium every 5 days, take the second harvest and third harvest of cell culture venom as the poisonous species for production;

[0041] Identification of cytotoxic species: The identification of cytotoxic species fully complies with the standard of attenuated strains of classical swine fever, which is safe for pigs and has no side effec...

Embodiment 3

[0049] (1) Selection of cells for seedling preparation: select porcine kidney (IBRS-2) cell line for use;

[0050] (2) Passage and culture of cells for seedling production: IBRS-2 cell line was digested and passed by EDTA-trypsin cell dispersion solution, and continued to culture at 37°C with cell growth medium. When a good monolayer was formed, it was used for continued passage or inoculation Virus;

[0051] (3) Propagation of cytotoxic species: use cell maintenance solution to make 0.5% virus suspension from fresh spleen venom, inoculate a single layer of well-growing IBRS-2 cell line, and place it at 37°C to continue culturing. Harvest and change the medium every 4 days, take the second harvest and third harvest of cell culture venom as the poisonous species for production;

[0052] Identification of cytotoxic species: The identification of cytotoxic species fully complies with the standard of attenuated strains of classical swine fever, which is safe for pigs and has no s...

Images