What antifungal to use in Tissue Culture??? - (Nov/22/2011 )

Hi,

Just wondering if anyone knows what antifungal I should use in tissue culture?? I am about to start an invassion assay looking at the invassion of Aspergillus fumigatus in epithelial cells. I need to use an antifungal that will kill the extracelluler fungal spores(conidia) but not destroy or lyse the epithelial cells. I want to look at conidia that were internalised by the epithelial cells and see if they grow! Any suggestions????? Any suggestions or advice would be great. Thanks

Hola, the most used antifungic for cell culture is amphotericin= Fungizone but I´m not sure that it serves for your purpose. Buena suerte

I don't know much about fungi but here are some of the questions I'd be asking (and someone may be able to answer these here)-

1. Is an anti fungal able to work only extracellularly and not be internalised by the cells, killing any intracellular fungi?
2. Won't your anti fungal kill any of the fungus that grows after invasion, which will affect your ability to measure this growth?
3. an anti fungal may have unwanted effects on your epithelial cells?

My approach would be to inoculate the cells with fungus, leave for defined infection period, then wash away any fungus that hadn't invaded (not sure what kind of wash buffer, you'd have to find out how the fungus adheres prior to invasion to decide which buffer was best) and then go from there.

I suppose it might be possible to have a short incubation in an anti fungal (long enough to kill any extracellular fungus but not affect your intracellular) and then replace with anti-fungal free media?

Sorry, that isn't much help probably!