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Serum-free medium and its application and a culture method of swine fever virus

A technology of serum-free medium and basal medium, applied in the field of swine fever virus culture and serum-free culture, can solve the problems of inability to directly learn from, and the culture conditions are very different, and achieve good amino acid metabolism, fast growth rate, Rapid proliferation effect

Active Publication Date: 2017-11-17
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Since the culture conditions required by different cells or viruses are very different, the inventors cannot directly learn from the composition of the existing serum-free medium, and must address this problem, re-study, and obtain suitable for ST cell culture, and CSFV Serum-free medium for proliferation

Method used

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  • Serum-free medium and its application and a culture method of swine fever virus
  • Serum-free medium and its application and a culture method of swine fever virus
  • Serum-free medium and its application and a culture method of swine fever virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0146] Dissolve the following substances respectively, mix each solution, and stir in the dark for 30 minutes until fully mixed, then add 800ml of ultra-clean pure water without pyrogen, stir and mix again, set the volume to 1000ml, and adjust the pH of the solution to 7.2. Added substances include: MEM basal medium 9.8g; sodium pyruvate 50-200mg; soybean protein hydrolyzate 0.1-2g; cholesterol 0.1-6mg; basic fibroblast growth factor 0.5-50mg; ;Insulin 5~50mg; L-alanine 2~30mg; L-valine 52~150mg; L-leucine 10~80mg; L-isoleucine 5~180mg; L-methionine 15 ~300mg; L-proline

[0147] 10~250mg; L-Phenylalanine 10~250mg; L-Tryptophan 5~300mg; L-Glycine

[0148] 0~50mg; L-serine 5~150mg; L-threonine 5~250mg; L-cysteine ​​5~250mg; L-asparagine 5~200mg; L-glutamine 250~400mg; L- Tyrosine 5~250mg; L-Aspartic Acid 2~80mg; L-Glutamic Acid 5~200mg; L-Lysine 6~100mg; L-Arginine

[0149] 5~300mg; L-histidine 5~300mg; Hydroxyproline 0~40mg; Myristic acid 0.01~0.3mg; Palmitic acid 0.01~0.3mg...

Embodiment 2

[0153] According to the mode in Example 1, prepare the first kind of target serum-free medium, wherein each substance is as shown in Table 1,

[0154] Table 1 Components of serum-free medium suitable for ST cell growth

[0155] MEM basal medium

[0156] L-Glycine

Embodiment 3

[0158] Dissolve the following substances respectively, mix each solution, and stir in the dark for 30 minutes until fully mixed, then add 800ml of ultra-clean pure water without pyrogen, stir and mix again, set the volume to 1000ml, and adjust the pH of the solution to 6.6. Added substances include: MEM basal medium 9.8g; sodium pyruvate 50-300mg; soybean protein hydrolyzate 0.1-2g; cholesterol 2-6mg; basic fibroblast growth factor 0.5-20mg; ;Insulin 10~50mg; L-alanine 10~100mg; L-valine

[0159] 2~35mg; L-methionine 10~30mg; L-proline 10~250mg; L-tryptophan 5~300mg; L-glycine 10~50mg; L-serine 5~150mg; L-cysteine Amino acid 5~250mg; L-Asparagine 5~200mg; L-Glutamine 100~200mg; L-Aspartic Acid 2~80mg; L-Glutamic Acid 5~200mg;

[0160] Hydroxyproline 10~60mg; Myristic acid 0.01~0.3mg; Palmitic acid 0.01~0.3mg; Palmitoleic acid 0.01~0.3mg; Stearic acid 0.01~0.3mg; Spermine 0.1~0.3ml; Spermidine 0.1~ 0.3ml; reduced glutathione 3~15mg; ethanolamine 1.5~3ml; β-mercaptoethanol 1~2...

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Abstract

The present invention relates to the field of biotechnology, in particular to the technical field of veterinary biological products engineering, more specifically to the serum-free culture and its application and the culture method of swine fever virus using these medium. The present invention uses MEM as the basic medium, through the study of the growth and proliferation characteristics of pig testicular cells and classical swine fever virus, and by adding different components in the basic medium, a series of serum-free medium for pig testicular cells and different The serum-free medium for the proliferation of classical swine fever virus in the growth stage not only avoids the risk of contamination caused by BVDV and its antibodies, but also enables ST cells to grow adherently, with rapid growth, complete cell surface, and plump individuals, which can be subcultured for a long time .

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to the technical field of veterinary biological products engineering, more specifically to the serum-free culture and its application and the culture method of swine fever virus using these medium. Background technique [0002] Classical swine fever (CSF) is a highly contagious disease caused by classical swine fever virus (CSFV), and is listed as a category A zoonotic disease by the World Organization for Animal Health. At present, the main means of controlling swine fever epidemic disease in my country is to inoculate the live vaccine of classical swine fever lanovirus (HCLV). The traditional production method of swine fever vaccine is to infect rabbits, and then collect spleen and peritoneal lymph to make tissue vaccines, or use primary bovine testicular cells to proliferate cell vaccines. However, the above two methods have been gradually replaced by the continuous passage ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12N7/00C12R1/93
Inventor 冯磊褚轩吴培培王伟峰侯继波华涛陈丽
Owner JIANGSU ACAD OF AGRI SCI
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