Schematic setup of
MAPC, splenocyte, and microglia/macrophage
interactions. (A) Isolated splenocytes are activated with 2 μg/ml of ConA. Two hours after isolation, MAPC were added at a ratio of 1:5 (MAPC:splenocyte) in co-culture or transwell cultures. 24 hours after the first dose of MAPCs, a second dose of MAPCs were added at the same ratio. 48 hours after the splenocyte harvest, supernatants were collected from each specific condition. (B) The supernatant was then used to stimulate LPS-activated microglia. (C) Microglia/macrophages were characterized via flow cytometry, immunostained or measured for proliferation using the MTS proliferation assay. LPS, lipopolysaccharide; MAPCs, multipotent adult progenitor cells; MTS.