I run a co-culture experiment with HeLa (mammalian) cells and bacteria (P. bivia, P. disiens, L. crispatus). Following a co-incubation period i want to measure the viability of the HeLa cells using an MTT assay which relies on mitochondrial produced products to detect viable cells. The problem is that bacteria make a homologous version of this mitochondria product and this can lead to false positives in the viability assay? I can wash some of the bacteria off using PBS or centrifugation methods, however some of the bacteria will remain inside of the Hela cells. If i administer a Penicillin/streptomycin (PEST) cocktail would this kill invaded bacterial cells??
Dear Nicholas,
Streptomycin is a broad spectrum antibiotic so it should be effective.
However, perhaps you could perform a dose dependent study on your bacteria with Pen/Strep to determine MIC? Then you could determine whether at this MIC there is any effect on your HeLa?
Hi,
I am not sure about the following statement that you made: "The problem is that bacteria make a homologous version of this mitochondria product and this can lead to false positives in the viability assay?"
If this is the problem, then would this be avoidable even after the clearance of the bacteria from the assay since I assume bacteria would produce these before you would want to kill the bacteria.
Debki, yes I know but if the bacterial cell is killed would these mitochondrial homologs degrade given a 24 hr incubation period with a PEST cocktail?
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