Role of soluble factors and three-dimensional culture in in vitro differentiation of intestinal macrophages

Figure 6 Mixed spheroids of MO and IEC or the control cell line. Mixed spheroids of the IEC lines HT-29 and WiDr or the control cell line J82 and MO were generated and cultured for seven days, disaggregated and examined by flow cytometry. A: There was a strong decrease of surviving MO/MAC (CD33+ cells) inside IEC spheroids compared to control spheroids over the seven-day culture period. In HT-29 spheroids the percentage decreased from 21.7% (24 h) to 1.4% (7 d). In WiDr spheroids a slighter decrease was observed with 14.6% MO/MAC after 24 h and 2.4% after seven days. In spheroids of the control cell line J82 no selection of a MO/MAC subpopulation could be observed. The number of MO/MAC inside the aggregates was nearly constant with 15.2% (24 h) and 13.2 (7 d); B: Addition of a blocking anti-Fas antibody 30 min before generation of the mixed spheroids did not change the results. In IEC-MCS a strong decrease of MO/MAC was observed (HT-29: 14.2% 24 h, 0.8% 7 d, WiDr: 20.1% 24 h, 2.3% 7 d). The MO/MAC number in control cell MCS was nearly constant with 13.9% (24 h) and 12.0% (7 d).