Serum-free culture media and application thereof as well as culture method of classical swine fever virus
A serum-free culture medium and swine fever virus technology, applied in the field of serum-free culture and swine fever virus culture, can solve the problems of inability to directly learn from and great differences in culture conditions, and achieve good amino acid metabolism, fast growth rate, Rapid proliferation effect
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[0145] Example 1
[0146] Dissolve the following substances separately, mix each solution, stir for 30 minutes in the dark, until fully mixed, add 800ml of ultra-pure water without heat source, stir and mix again, dilute to 1000ml, and adjust the pH of the solution to 7.2. The added substances include: MEM basic medium 9.8g; sodium pyruvate 50~200mg; soybean protein hydrolysate 0.1~2g; cholesterol 0.1~6mg; basic fibroblast growth factor 0.5~50mg; inhibin B 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 ...
Example Embodiment
[0152] Example 2
[0153] According to the method in Example 1, the first target serum-free medium was prepared, and the substances are as shown in Table 1.
[0154] Table 1 Serum-free medium components suitable for ST cell growth
[0155] MEM basal medium 9.8g / L Sodium pyruvate 200mg / L Soy protein hydrolysate 2g / L cholesterol 2.5mg / L Basic fibroblast growth factor 25mg / L Inhibin B 25mg / L insulin 25mg / L L-alanine 20mg / L L-valine 150mg / L L-Leucine 45mg / L L-Isoleucine 58mg / L L-methionine 150mg / L L-Proline 150mg / L L-phenylalanine 150mg / L L-tryptophan 50mg / L
[0156] L-glycine 10mg / L L-serine 55mg / L L-threonine 55mg / L L-cysteine 55mg / L L-Asparagine 50mg / L L-glutamine 250mg / L L-tyrosine 55mg / L L-Aspartic Acid 20mg / L L-glutamic acid 50mg / L L-Lysine 60mg / L L-Arginine 50mg / L L-histidine 50mg / L Hydroxyproline 40mg / L Myristic acid 0.1mg / L Palmitic acid 0.1mg / L Palmitoleic acid 0.1mg / L Stearic acid 0.1mg / L Spermine 0.1ml / L Spermidine 0.1ml / L Reduced glut...
Example Embodiment
[0157] Example 3
[0158] Dissolve the following substances separately, mix each solution, stir for 30 minutes in the dark, until fully mixed, add 800ml of super-clean pure water without heat source, stir and mix again, dilute to 1000ml, and adjust the pH of the solution to 6.6. The added substances include: MEM basic medium 9.8g; sodium pyruvate 50~300mg; soybean protein hydrolysate 0.1~2g; cholesterol 2~6mg; basic fibroblast growth factor 0.5~20mg; inhibin B 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 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-me...
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