Hello,
The biofilm that I am culturing from this strain is too flimsy, such as when I wash the biofilm with PBS it breaks and resuspends even at a very gentle wash.
I have been growing this strain in the following conditions:
Liquid culture: 24 hours 37C with TSB, with or without 1% glucose at 250 RPM or stationary.
Biofilm formation: 24 hours 37C with TSB, with or without 1% glucose on a 96 well Polystyrene tissue culture plate (GenClone Cat# 25-109).
This is also what I can find in publications.
I am planning to try different media, sugars, concentrations, and plate combinations.
However, does anyone have any insight for specifics I should attempt to get a formidable biofilm?
Thanks!
Hi,
Adding some 2% NaCl can make staphylococcal biofilms a bit more robust. Another option is to add blood plasma. S. aureus can convert fibrinogen from plasma into fibrin and use it as a component of the biofilm matrix (so it is important to use plasma, not serum!). In our hands it makes staphylococcal biofilms much more resilient. However, it also changes other properties of the biofilms, particularly antibiotic resistance. We use 1.5% heparinized rat plasma inactivated at 56°C for 30 min, but the concentrations cited in the literature can be up to 50%.
A couple papers for more info:
Article An improved medium for growing Staphylococcus aureus biofilm
Article Morphological and Biological Characteristics of Staphylococc...
https://pubmed.ncbi.nlm.nih.gov/25034276/
https://pubmed.ncbi.nlm.nih.gov/26044292/
https://pubmed.ncbi.nlm.nih.gov/26136471/
Dear Oded,
I work with Pseudomonas app. biofilms where we also get weak or poorly-attached biofilms - as well as some really strong ones occasionally. Although I appreciated the fact that you may have a problem growing USA300-0114 biofilms as well as others may have reported for the same or similar strains, there is the theoretical question of why this needs to be done; i.e., strains may produce weaker or stronger biofilms under different conditions, and all of these are relevant unless you are simply wanting a uniform, reproducible 'in vitro' biofilm for study!
Andrew
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