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Title: In vitro and in vivo generation and characterization of Pseudomonas aeruginosa biofilm–dispersed cells via c-di-GMP manipulation
Authors: Chua, Song Lin
Hultqvist, Louise D.
Yuan, Mingjun
Rybtke, Morten
Nielsen, Thomas E.
Givskov, Michael
Tolker-Nielsen, Tim
Yang, Liang
Keywords: Biofilm dispersal
Pseudomonas aeruginosa
Issue Date: 2015
Source: Chua, S. L., Hultqvist, L. D., Yuan, M., Rybtke, M., Nielsen, T. E., Givskov, M., et al. (2015). In vitro and in vivo generation and characterization of Pseudomonas aeruginosa biofilm–dispersed cells via c-di-GMP manipulation. Nature Protocols, 10(8), 1165-1180.
Series/Report no.: Nature Protocols
Abstract: Bis-(3′-5′)-cyclic dimeric guanosine monophosphate (c-di-GMP) is a global secondary bacterial messenger that controls the formation of drug-resistant multicellular biofilms. Lowering the intracellular c-di-GMP content can disperse biofilms, and it is proposed as a biofilm eradication strategy. However, freshly dispersed biofilm cells exhibit a physiology distinct from biofilm and planktonic cells, and they might have a clinically relevant role in infections. Here we present in vitro and in vivo protocols for the generation and characterization of dispersed cells from Pseudomonas aeruginosa biofilms by reducing the intracellular c-di-GMP content through modulation of phosphodiesterases (PDEs). Unlike conventional protocols that demonstrate biofilm dispersal by biomass quantification, our protocols enable physiological characterization of the dispersed cells. Biomarkers of dispersed cells are identified and quantified, serving as potential targets for treating the dispersed cells. The in vitro protocol can be completed within 4 d, whereas the in vivo protocol requires 7 d.
ISSN: 1754-2189
DOI: 10.1038/nprot.2015.067
Rights: © 2015 Nature America, Inc. This is the author created version of a work that has been peer reviewed and accepted for publication by Nature Protocols, Nature America, Inc. It incorporates referee’s comments but changes resulting from the publishing process, such as copyediting, structural formatting, may not be reflected in this document. The published version is available at: [].
Fulltext Permission: open
Fulltext Availability: With Fulltext
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