Higher production of rabies virus in serum-free medium cell cultures on microcarriers.
Rabies virus suspensions were obtained from VERO cells cultivated on solid microcarriers in a bioreactor after infection with the Pasteur rabies virus strain (PV). Virus production-serum free medium (VP-SFM) or Leibovitz 15 (L15) medium supplemented or not with fetal calf serum (FCS) were used to cultivate the VERO cells, before and after virus infection. The cell growth was shown to reach higher densities (1.6 x 10(6) cellsmol(-l)), when VP-SFM supplemented with 1% of FCS was used during the cell growth phase of culture, and then replaced by VP-SFM alone for the virus multiplication phase. In the cultures performed from the beginning with VP-SFM, lower densities accompanied by an altered cell morphology and detachment from the microcarriers were always observed. In rabies virus infected cultures, kinetic studies showed that higher virus yields (10(4.7) FFD(50) per 0.05 ml) were always obtained in cultures performed initially on VP-SFM supplemented with 1% FCS and after infection on VP-SFM alone. In agreement with that, rabies virus production, as measured by the average of virus titers in harvests obtained at different times after infection were shown to be 5.5 times higher in the cell cultures using initially VP-SFM+1%FCS and, following infection, VP-SFM alone. Besides the advantages of using media with a well-controlled composition, these data indicate the usefulness of serum free media also in terms of virus productivity.
Frazzati-Gallina
N M
NM
Laboratório de Raiva, Instituto Butantan, Av. Vital Brasil 1500, 05503-900 Sao Paulo, Brazil.
Paoli
R L
RL
Mourão-Fuches
R M
RM
Jorge
S A
SA
Pereira
C A
CA
D000818:Animals; D019149:Bioreactors; D002455:Cell Division; D002478:Cells, Cultured / Q000821:virology; D016895:Culture Media, Serum-Free; D011820:Rabies virus / Q000254:growth & development*; D014776:Virus Cultivation / Q000295:instrumentation / Q000379:methods
D016895:Culture Media, Serum-Free