i am doing THP-1 suspension culture and using cefotaxim as a antibiotic but after 4 passag my cell count was decline, so is it becuase cefotaxim my cell is decline
Commonly used Antibiotic in for THP-1 growth Medium is 1% Pen/strep. I Have used 25 ug/mL Gentamicin as well, with no apparent effect on cell growth rate/viability.
If Cefotaxime is the only antibiotic available to you right now, I would suggest to do growth curves for THP-1 cells under different concentrations of Cefotaxime(make sure to include negative control for comparison). This will help to see if Cefotaxime, at chosen concentration, is compromising THP-1 cell culture viability.
As a side note, THP-1 cells are the most sensitive cell line I’ve ever worked with. I would suggest to maintain conditions listed below to make sure your THP-1 cells are happy
· Complete Medium: RPMI-1640+10% FBS( heat inactivated)+2-Mercaptoethanol( 0.05 mM)
· Maintain seeding density in your culture flask between 5 x10^5 to 1 x10^6 cells/mL.
· Do not passage your cells line more than 3 months( or approximately 12 passages)
· During centrifugation, spin down between 125-300g for 5-10 minutes.
Hello Yash,
You need to obtain some information on cefotaxime before you use it in culture, as rightly stated by Nino Mtchedlidze . Generally, Penicillin/Streptomycin mixture (PS) and Gentamicin are widely used as standard antibiotics in cell culture. The combination of Penicillin at (100 U/mL) and streptomycin at (100 µg/mL) are used or Gentamycin at a standard concentration of (50 µg/mL) is used.
But why do you want to use antibiotics in culture? If you strictly maintain sterile environment in cell culture laboratory, you may avoid using antibiotics in cell culture. Antibiotics are biologically and pharmacologically active. In addition to the emergence of antibiotic-resistant bacteria, antibiotics could have multi-level effects on mammalian cells and may react with any agent or substance in culture media. So, antibiotic-free culture media are recommended to ensure the reliability and reproducibility of mammalian cell culture system-derived scientific findings.
Best.
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