Varicella attenuation live vaccine
A technology of attenuated live vaccine, varicella, applied in the field of medical microbiology
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Embodiment 1
[0040] A live attenuated varicella vaccine, prepared as follows:
[0041] (1) MRC-5 cells were continuously passaged for 20 generations in cell flasks;
[0042] (2) After the cells grow to a certain scale, the virus seed is inoculated into the cells, and two consecutive infections are carried out, and the M.O.I is respectively 0.001 and 0.01 to expand the harvest of the virus liquid;
[0043] (3) adding MEM culture solution containing 2% fetal bovine serum, and culturing at 35°C;
[0044] (4) When the lesion degree of MRC-5 cells reaches ++~+++, pour out the culture medium, wash the cell surface with PBS washing solution, and wash away the bovine serum;
[0045] (5) Add 0.05% EDTA solution to elute the infected cells, collect the cell suspension in a centrifuge tube, centrifuge at 1800 rpm for 5 minutes at 4°C, discard the supernatant, add the vaccine solution containing a stabilizer, mix well, and collect , aspirated for single-bottle sterility testing, and stored at -80°C;...
Embodiment 2
[0052] A live attenuated varicella vaccine, prepared as follows:
[0053] (1) MRC-5 cells are continuously passaged in cell flasks for no more than 25 generations;
[0054] (2) After the cells grow to a certain scale, the virus seed is inoculated into the cells, and two consecutive infections are carried out, and the M.O.I is respectively 0.0015 and 0.02, so as to expand the harvest of the virus liquid;
[0055] (3) Add MEM culture solution containing 2.5% fetal bovine serum, and culture at 36°C;
[0056] (4) When the lesion degree of MRC-5 cells reaches ++~+++, pour out the culture medium, wash the cell surface with PBS washing solution, and wash away the bovine serum;
[0057] (5) Add 0.05% EDTA solution to elute the infected cells, collect the cell suspension in a centrifuge tube, centrifuge at 1800 rpm for 5 minutes at 6°C, discard the supernatant, add the vaccine solution containing a stabilizer, mix well, and collect , aspirated for single-bottle sterility testing, and...
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