Protein response of insect cells to bioreactor environmental stresses.
Protein expression of Spodoptera frugiperda (Sf9) insect cells was characterized upon exposure to environmental stresses typically present in bioreactors including heat shock, oxygen deprivation, shear stress, change of pH, and salinity or ethanol shock. This study fills the void in knowledge as to how bioreactor hydrodynamics, anoxia, small changes in pH as well as salinity alterations due to pH control or exposure to ethanol used in asepsis treatments affect protein expression in Sf9 cells. Heat shock at 43 degrees C induced proteins at 83 kDa, 68-78 kDa and six small heat shock proteins (hsps) at 23-15.5 kDa. Anaerobic conditions in CO2 atmosphere reduced significantly the normal protein synthesis and induced a small subset of heat shock proteins at 70 kDa. Oxygen deprivation in nitrogen atmosphere transiently induces the 70 kDa proteins and had minor effects on the normal protein synthesis. Exposure to increased salinity or ethanol concentration failed to trigger the stress response, but may extensively inhibit the induction of normal proteins even though there was a negligible change in cell viability. Shear stress that had a major reducing effect on cell viability did not change the protein synthesis profile of Sf9 cells. Both long and short term exposures to small pH changes had negligible effects on protein synthesis.
Department of Chemical Technology, Helsinki University of Technology, P.O. Box 6100, FIN-02150 HUT, Finland.
Anne
A
Huhtala
Pekka
P
Linko
Raj
R
Mutharasan
D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D004777:Environment; D000431:Ethanol / Q000494:pharmacology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology*; D018869:Heat-Shock Response / Q000187:drug effects / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D019476:Insect Proteins / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology*; D010100:Oxygen / Q000378:metabolism; D018411:Spodoptera / Q000187:drug effects / Q000378:metabolism*; D013696:Temperature
D019476:Insect Proteins; D000431:Ethanol; D010100:Oxygen