Relationship between oxygen uptake rate and time of infection of Sf9 insect cells infected with a recombinant baculovirus.
Oxygen uptake rates (OUR) of Sf9 insect cells propagated in a serum-free medium (SF900II, Gibco) and of cells infected with a recombinant AcNPV were investigated before and after infection in a laboratory-scale bioreactor. The volumetric OURs of uninfected and exponentially growing cells were found to be proportional to the cell density. For infected cultures, the specific OUR of cells increased immediately after addition of virus and a maximum of 1.3 times the value of uninfected cells was noted for all the cultures between 8 to 30 hours post infection, which coincides with the period at which most viral replication and the majority of DNA synthesis takes place. It was observed that the rate of rise in the specific OUR decreased as the cell density at the time of infection increased, which meant that the later the infection, the later the maximum sOUR was observed. We therefore suggest that OUR measurement can be used to reflect the efficiency of a batch infection. Carbohydrate and amino acid consumption rates from an infected run were analysed in an effort to identify substrate(s) that may be used at increased rates to fuel the rise in oxygen demand observed early in the infection cycle. No observable rise in the consumption rates of glucose or glutamine, which are the major energy sources for animal cells, were seen after infection but an increase in the consumption rates of some amino acids suggests that infected Sf9 cells may utilise amino acids at an enhanced rate for energy post infection.
Wong
T K
TK
Department of Chemical Engineering, University of Queensland, Australia.
Nielsen
L K
LK
Greenfield
P F
PF
Reid
S
S
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D016367:Baculoviridae; D001709:Biotechnology / Q000295:instrumentation / Q000379:methods; D050260:Carbohydrate Metabolism; D002460:Cell Line; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods; D005822:Genetic Vectors; D007700:Kinetics; D008962:Models, Theoretical; D010101:Oxygen Consumption; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis*; D011995:Recombination, Genetic; D018411:Spodoptera; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D001616:beta-Galactosidase / Q000032:analysis / Q000096:biosynthesis*
D000596:Amino Acids; D011994:Recombinant Proteins; D001616:beta-Galactosidase