Effect of MAPC:Splenocyte co-culture
supernatant on stimulated microglial
immunophenotype. (A) Samples of microglial cultures were first gated on forward- and side-scatter characteristics to exclude debris, electronic noise, and aggregates (not shown). Resulting populations were then gated to exclude a small number of artifacts displaying extremely high signal for either CD206-AF488 or CD86-PE. Rather than divide this microglia population further by imposing strict boundaries on a continuous expression pattern, mean fluorescence intensity (MFI, listed in each box) of the entire remaining population was then determined for each marker and the CD206/CD86 MFI ratio (M2:M1 ratio) calculated. (B) There was a significant increase in the M2:M1 ratio when the microglia received the supernatant derived from MAPC:splenocyte co-culture plus LPS when compared to microglia which only received LPS alone, or supernatant derived from MAPC:splenocyte transwell cultures plus LPS. ** represents P <0.01. LPS, lipopolysaccharide; MAPC, multipotent adult progenitor cell.