Quantitation of interaction of lipids with polymer surfaces in cell culture.
As cell culture medium development efforts have progressed towards leaner, serum-free, and chemically defined formulations, it has become increasingly important to ensure that the appropriate concentrations of all nutrients are maintained and delivered at point of use. In light of concurrent efforts to progress to disposable polymeric storage and culture platforms, the characterization and control of medium component interactions with container surfaces can be a key issue in ensuring consistent delivery of these medium formulations. These studies characterize the interactions of lipids with culture surfaces typically encountered in the bioprocess industry using model systems. The extent and kinetics of lipid association with polymeric surfaces were determined using radio-labeled linoleic acid and cholesterol. The effect of methyl-beta-cyclodextrin, a component commonly used to solubilize lipids in culture media, on association kinetics was also examined. In addition, loss of lipids across a sterilizing membrane filter was quantified. We find that there is potential for significant loss of hydrophobic components due to non-specific binding to surfaces at timescales relevant to a typical cell culture process. The extent of loss is dependent on the nature of the hydrophobic component as well as the type of surface. These studies highlight the potential of the extracellular environment to modify medium composition and also emphasize the importance of medium formulation strategies, including those used in the delivery of hydrophobic components. It is noted, however, that the level of loss is very dependent on the specific system including the composition of the culture medium used.
Altaras
Gina M
GM
Fermentation and Cell Culture, Bioprocess R&D, Merck Research Laboratories, Merck & Co., Inc., WP17-201, 770 Sumneytown Pike, West Point, PA 19486, USA.
Eklund
Carrie
C
Ranucci
Colette
C
Maheshwari
Gargi
G
D018929:Cell Culture Techniques; D003470:Culture Media / Q000737:chemistry*; D004347:Drug Interactions; D005227:Fatty Acids / Q000378:metabolism; D008055:Lipids / Q000032:analysis*; D008954:Models, Biological; D011108:Polymers / Q000737:chemistry / Q000378:metabolism*
D003470:Culture Media; D005227:Fatty Acids; D008055:Lipids; D011108:Polymers