I need a protocol to generate persistent cells of S. epidermidis because I want to test my agent only against to persiter cells of s epidermidis.
I think maybe I could growth a biofilm and treat with vancomicin ?
thanks
Dear Lydia, maybe the following paper can help you.
Best regards.
https://www.researchgate.net/publication/258523357_Activated_ClpP_kills_persisters_and_eradicates_a_chronic_biofilm_infection
Article Activated ClpP kills persisters and eradicates a chronic bio...
There is a chapter related to your question in "Biofilm-based Health care associated
Infections, Volume II" Advances in Experimental Medicine and Biology 831, Persister Cells in Biofilm Associated Infections
by Brian P. Conlon, Sarah E. Rowe, and Kim Lewis
We have successfully followed this protocol: Grow the biofilms on polystyrene support for 72 h. Carefully aspirate media and planktonic cells. Add ciprofloxacin (50 μg/mL) (for S. epidermidis) or Vancomycin (20 μg/mL) (for MRSA) in ½ x TSB and incubate for 24 h. Scrape the biofilm with a sterile cell scraper and wash the suspended cells in saline. You can look for persisters in this using confocal microscopy by using the LIVE/DEAD® BacLight™ Bacterial Viability Kit or use an esterase dye like calcein. You can also use FACS Sorter to isolate pure persisters from this suspension. If you don’t have these systems, simply follow up the scraping with serial dilution and plating on TSA containing 1% MgCl2 , to inactivate ciprofloxacin. The colonies will usually be small. Best of luck!