Pluronic enhances the robustness and reduces the cell attachment of mammalian cells.
The addition of the non-ionic surfactant, Pluronic F-68, to serum-free CHO cultures causes multi-functional effects that enhance cell yield in agitated cultures and reduce cell adhesion in stationary cultures. Three independent CHO cell lines were subjected to high liquid shear in assay systems that either included or excluded a liquid-gas interface. In the absence of Pluronic, there was a loss in cell viability in either assay system, although there was an intrinsic variability in sensitivity of the cell lines to shear damage. Supplementation with Pluronic prevented loss of cell viability, indicating protection in either a gas sparged or bubble-free environment. However, we found no evidence of long-term protection of cells once Pluronic was removed. Pluronic was capable of repairing trypsin-damaged cells as evidenced by enhanced growth, reduced membrane porosity, and improved robustness under liquid shear. The proportion of adherent cells was reduced to a minimal level by the presence of Pluronic although its effect was rapidly reversible with a high proportion (70%) of adherent cells observed within a few culture passages of its removal. The observed effects of Pluronic on these cultures are compatible with a mechanism in which the polymer forms a protective layer on the cell membrane, which has a significantly lower hydrophobicity.
Tharmalingam
T
T
Department of Microbiology, University of Manitoba, Manitoba, Winnipeg, Canada R3T 2N2.
Ghebeh
H
H
Wuerz
T
T
Butler
M
M
D000818:Animals; D019149:Bioreactors; D016466:CHO Cells; D002448:Cell Adhesion / Q000187:drug effects*; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002463:Cell Membrane Permeability; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media; D057927:Hydrophobic and Hydrophilic Interactions; D020442:Poloxamer / Q000494:pharmacology*; D033081:Shear Strength; D013314:Stress, Mechanical; D013501:Surface-Active Agents / Q000494:pharmacology*; D014343:Trypan Blue / Q000493:pharmacokinetics; D014357:Trypsin / Q000494:pharmacology; D014783:Viscosity
D003470:Culture Media; D013501:Surface-Active Agents; D020442:Poloxamer; D014357:Trypsin; D014343:Trypan Blue