Virus Production Technology Research Vero Cells With Low Serum Microcarriers In Suspension Culture Model Optimization And Oncolytic Herpes Simplex

Cancer has been become the first fatal human disease now, using of Vero cells for recombinant HSV-Ⅱ production, which has a good of Lysogenic, targeting and safety, having a huge importance influence for large scale industrial production.In this work, low serum medium VD-LSM containing1%NBCS was designed base on DMEM/F121:1medium. The maximum viable cell density reached about1.75×106cells/ml in T-25flask and remained stable after successive passages for30s.On the basis of the metabolic analysis of Vero cells in in batch culture in1.5L bioreactor, the maximum viable cell density reached about4.06×106cells/ml in fed-batch culture developed, higher than the batch culture of2.82×106cells/ml.The bead-to-bead transfer on Cytodex1of Vero cells mainly rely on the formation of cell bridges between old and fresh microcarriers, the research about this was development. The optimal condition of transfer were about30cells/MC inoculated, cultured for60h when the cells density reached about the same density with the initial inoculated, the static time/stiring time was42min/3min. It can be achieved for four times successive transfer for amplification by the1:4(V:V), cells density reach up to6.0×106cells/ml.Vero cells culture on Cytodex1was carried out for the scale up production of HSV-Ⅱ virus in a5L bioreactor on the basis of above work. The optimal culture conditions were:low-serum medium VD-LSM with1%NBCS, MOI0.03, TOI36h (after the inoculated), pH7.14, T32℃, received after cultured for96h. The process of HSV-Ⅱ virus production was developed, the titer is reached up to5.6units/100μl above.