查看文章

MATERIALS AND METHODS

Organisms. C. albicans 3153A was used in the course of this study, and was stored on Sabouraud dextrose slopes (BBL, Cockeysville, Md) at–70 C. C. albicans 3153A was propagated in yeast peptone dextrose (YPD) medium (1% w/v yeast extract, 2% w/v peptone, 2% w/v dextrose [US Biological, USA]). Batches of medium (20 ml) were inoculated from YPD agar plates containing freshly grown C. albicans, and incubated overnight in an orbital shaker at 30 C. C. albicans 3153A grew in the budding-yeast phase under these conditions. Cells were harvested and washed in sterile phosphate buffered saline (PBS: 10 mM phosphate buffer, 2.7 mM potassium chloride, 137 mM sodium chloride, pH 7.4 [Sigma, USA]). Cells were resuspended in RPMI-1640 supplemented with L-glutamine and buffered with morpholinepropanesulfonic acid (MOPS)(Angus Buffers and Chemicals, USA) and adjusted to the desired cellular density by counting in a haematocytometer (see below).