Study On Live Attenuated Varicella Cell Factory Technology

Objective The stock solution and vaccine were produced according to optimized parameter,which virus was cultured in the cell factory,so the technology was prelimitary accomplished, and the feasibility and stability were assessed.Method Human dipioid cells of appropriate generation were cultured in the cell factory. When densified monolayer cells were cultured, Oka strain viruses were adsorbed on the monolayer cells. When cell pathology effect(CPE) come to 5%-10%,the monolayer cells were washed three times to wipe serum. When CPE come to70%,the cells were dislodged with EDTA,collected by centrifugation and stored-70℃. Infected cells were disrupted,clarified and and lyophilized, so vaccines were assessed according to 《Chinese Pharmacopoeia(3rd edition) 》and 《the Regulation of Live,Herpes-zoster virus Vaccine Production and Test》.Result Human dipioid cells of 36 generation were transferred to 4 layers cell factory(CF4) at ratio 1:2.When MOI is 0.01-0.001,CPE is optical.When CPE reach to5%-10%, 300-400 ml PBS was added to CF4 per layer, and when CPE is peak period,50 ml 0.05%EDTA was applied to CF4.Virus harvest was collected by centrifugation at 1500rpm/5min in 4 ℃.The stock solution and vaccine were qualified.Conclution The key parameters that live,attenuated varicella vaccines were produced in CF4 were optimized; the production technology was established successfully; Preparation of vaccines were qualified;feasibility and stability of technology were verified.