The effect of spent medium recycle on cell proliferation, metabolism and baculovirus production by the lepidopteran Se301 cell line infected at very low MOI.
The aim of this paper was to study the effect of spent medium recycle on Spodoptera exigua Se301 cell line proliferation, metabolism, and baculovirus production when grown in batch suspension cultures in Ex-Cell 420 serum-free medium. The results showed that the recycle of 20% of spent medium from a culture in mid-exponential growth phase improved growth relative to a control culture grown in fresh medium. Although both glucose and glutamine were still present at the end of the growth phase, glutamate was always completely exhausted. The pattern of the specific glucose and lactate consumption and production rates, as well as the specific glutamine and glutamate consumption rates, suggests a metabolic shift at spent medium recycle values of over 60%, with a decrease in the efficiency of glucose utilization and an increase in glutamate consumption to fuel energy metabolism. Baculovirus infection provoked a change in the metabolic pattern of Se301 cells, although a beneficial effect of spent medium recycle was also observed. Both growth rate and maximum viable cell density decreased relative to uninfected cultures. The efficiency of glucose utilization was dramatically reduced in those cultures containing the lowest percentages of spent medium, whereas glutamine and glutamate consumption was modulated, thereby suggesting that infected cells were devoted to virus replication, retaining their ability to incorporate the nutrients required to support viral replication. Recycle of 20% of spent medium increased baculovirus production by around 90%, thus showing the link between cell growth and baculovirus production.
Department of Chemical Engineering, University of Almería, 04120 Almería, Spain.
Alba
A
Beas-Catena
Asterio
A
Sánchez-Mirón
Francisco
F
Garía-Camacho
Antonio
A
Contreras-Gómez
Emilio
E
Molina-Grima
D000818:Animals; D016367:Baculoviridae / Q000254:growth & development*; D002460:Cell Line; D049109:Cell Proliferation; D002470:Cell Survival; D003470:Culture Media / Q000737:chemistry*; D005947:Glucose / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism; D007773:Lactates / Q000378:metabolism; D018411:Spodoptera
D003470:Culture Media; D007773:Lactates; D005973:Glutamine; D018698:Glutamic Acid; D005947:Glucose