The Availability of Large Scale VERO Cell Serial Subcultivation in Bioreactor

Objective:To develop a novel process for scaling up VERO cell culture by serial subcultivation on microcarrier from 5-liter bioreactor to 30-liter bioreactor.Method:VERO cells were grown on Cytodex-I in 5-liter bioreactor containing 10 g/L of microcarrier.After an initial 96 h perfusion cell culture,Pre-washed the confluent microcarriers,trypsinized and inoculated from 5-liter to 30-liter bioreactor containing 10 g/L of microcarrier,At the different period of cell culture in 30-liter bioreactor,used the uncleus extrusion method to determine cell number,the content of lactic acid and glucose were made an assay in 30-liter bioreactor.Result:After an initial 96 h perfusion cell culture in 5-liter bioreactor,the ratio of confluent microcarrier reached 95 percent,the density of cultured cells reached 7.81×106cells/mL ;the ratio of havesting cell from 5-liter bioreactor microcarriers reached 32.3 percent,and after 144 h cell culture in 30-liter bioreactor,the ratio of confluent microcarrier reached 90 percent,the density of cultured cells reached 9.19×106cells/mL.Conclusion:The process of serial subcultivation of VERO cell on microcarrier is most cost-effective means for inoculating the following larger vessel.