Dendritic cells used for analysis of microrganism survival following uptake

After 6 hour of stimulation, DCs were collected, washed 3 times with PBS, treated with zymolyase, washed twice and cells, lysated with a hypotonic solution (KCl 0.05%), were plated on YPD. Survival of yeast cells, spores or hyphae after uptake was reported as percentage of colony forming units after 3 days relative to the total number of cells growing in the absence of DCs exposure. To assess the importance of ROS production in DC killing ability, DPI (10 microM) was added 30 minutes before stimulation and survival of microorganisms was assessed using the same method. When evaluating survival after exposure, the possible effect of DMSO and DPI on the stimuli was taken into account.

created over 14 years ago (21 January 2010)    last modified over 13 years ago (28 September 2011)   [ RDF ]   [ RelFinder ]