A method for constructing a rhino-horned catfish heart cell line
A rhinoceros horn catfish and construction method technology, applied in artificial cell constructs, microbial-based methods, animal cells, etc., can solve problems such as no similar reports, achieve repeatability, low pollution rate, and simple and easy operation line effect
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Embodiment 1
[0026] 1. Prepare PBS disinfectant and cell culture medium
[0027] PBS disinfectant: Add antibiotics to PBS so that the concentration of penicillin is 100IU / ml, the concentration of streptomycin is 100μg / ml, the concentration of streptomycin sulfate is 40μg / ml, and the concentration of amphotericin B is 40μg / ml.
[0028] Basal culture medium: add glutamine and fetal bovine serum to the M199 medium, so that the concentration of glutamine is 300 μg / ml, and the fetal bovine serum accounts for 15% of the total volume.
[0029] Primary culture fluid: Add glutamine, fetal bovine serum and antibiotics to the M199 culture fluid, so that the concentration of glutamine is 600 μg / ml, fetal bovine serum accounts for 25% of the total volume, and the concentration of penicillin is 100 IU / ml. The streptomycin concentration was 100 μg / ml, the streptomycin sulfate concentration was 40 μg / ml, and the amphotericin B concentration was 40 μg / ml.
[0030] Subculture medium: Add glutamine, fetal b...
Embodiment 2
[0046] 1. Prepare PBS disinfectant and cell culture medium
[0047] PBS disinfectant: Add antibiotics to PBS so that the concentration of penicillin is 300IU / ml, the concentration of streptomycin is 300μg / ml, the concentration of streptomycin sulfate is 20μg / ml, and the concentration of amphotericin B is 20μg / ml.
[0048] Basal culture medium: add glutamine and fetal bovine serum to the M199 medium, so that the concentration of glutamine is 400 μg / ml, and the fetal bovine serum accounts for 10% of the total volume.
[0049] Primary culture solution: Add glutamine, fetal bovine serum and antibiotics to the M199 culture medium, so that the concentration of glutamine is 800 μg / ml, fetal bovine serum accounts for 15% of the total volume, and the concentration of penicillin is 200 IU / ml. The streptomycin concentration was 30 μg / ml, the streptomycin sulfate concentration was 20 μg / ml, and the amphotericin B concentration was 20 μg / ml.
[0050] Subculture medium: Add glutamine, feta...
Embodiment 3
[0061] 1. Prepare PBS disinfectant and cell culture medium
[0062] PBS disinfectant: Add antibiotics to PBS so that the concentration of penicillin is 300IU / ml, the concentration of streptomycin is 300μg / ml, the concentration of streptomycin sulfate is 20μg / ml, and the concentration of amphotericin B is 20μg / ml.
[0063] Basal culture medium: add glutamine and fetal bovine serum to the M199 medium, so that the concentration of glutamine is 400 μg / ml, and the fetal bovine serum accounts for 10% of the total volume.
[0064] Primary culture solution: Add glutamine, fetal bovine serum and antibiotics to the M199 culture medium, so that the concentration of glutamine is 800 μg / ml, fetal bovine serum accounts for 15% of the total volume, and the concentration of penicillin is 200 IU / ml. The streptomycin concentration was 30 μg / ml, the streptomycin sulfate concentration was 20 μg / ml, and the amphotericin B concentration was 20 μg / ml.
[0065] Subculture medium: Add glutamine, feta...
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