Mosquito and mammalian cells grown on microcarriers for four-serotype dengue virus production: variations in virus titer, plaque morphology, and replication rate.
Dengue (DEN) viruses consisting of four distinct serotypes cause diseases such as dengue fever, dengue hemorrhagic fever, and dengue shock syndrome in humans. Most of the dengue viruses can be effectively propagated in some mosquito and mammalian cell lines. In this study, we applied microcarrier cell culture technology to study two relevant aspects involving dengue virus, one on biotechnology of cell growth and virus production, and the other on virus biology concerning genetic variation of a virus population. We investigated the growth of C6/36 mosquito cells and Vero cells grown on Cytodex 1 microcarriers. High-titer DEN virus production can be achieved in C6/36 and Vero cells infected at low cell inoculation density, in the lag-phase cell stage, and at low multiplicity of infection (MOI). The maximum titers produced for DEN-1, DEN-3, and DEN-4 viruses were approximately 10- to 10,000-fold lower than for DEN-2 virus produced in C6/36 and Vero cells grown on microcarriers. The DEN-2 virus produced in C6/36 cells displayed far more extensive plaque heterogeneity than in Vero cells. Microcarrier C6/36 mosquito cell culture appeared to be the most effective system for four-serotype DEN virus production. Interestingly, some selected variants of DEN virus may outgrow in Vero cells when using a T-flask culture. These results may provide useful information for DEN vaccine development.
Liu
Chia-Chyi
CC
Institute of Biotechnology and Department of Life Science, National Tsing Hua University, Hsinchu 30013, Taiwan.
Wu
Suh-Chin
SC
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division; D002522:Chlorocebus aethiops; D018920:Coculture Techniques / Q000379:methods*; D009033:Culicidae / Q000821:virology*; D003716:Dengue Virus / Q000145:classification / Q000254:growth & development* / Q000302:isolation & purification / Q000648:ultrastructure*; D012703:Serotyping / Q000379:methods; D014709:Vero Cells; D010948:Viral Plaque Assay / Q000379:methods*; D014776:Virus Cultivation / Q000379:methods*; D014779:Virus Replication / Q000502:physiology