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

Figure 2 Flow cytometrical quantification of MO/MAC antigen expression after seven days of culture in IEC conditioned medium. A: Ninety-two percent of CD33+ cells (MO/MAC) showed expression of CD14 in MO cultured in unconditioned control medium (RPMI) without FCS supplemented with 2% human AB serum for seven days; B: Ninety-five percent of CD33+ cells (MO/MAC) were CD14-positiv in MO cultured in WiDr conditioned medium without FCS supplemented with 2% human AB serum for seven days; C: Ninety-two percent of CD33+ cells (MO/MAC) were CD14-positive in MO cultured in J82 conditioned medium (control cell line of non-intestinal origin) without FCS supplemented with 2% human AB serum for seven days; D: Ninety-four percent of CD33+ cells (MO/MAC) showed expression of CD14 in MO cultured in unconditioned control medium (DMEM) without FCS supplemented with 2% human AB serum for seven days; E: Ninety-eight percent of CD33+ cells (MO/MAC) were CD14-positive in MO cultured in HT-29 conditioned medium without FCS supplemented with 2% human AB serum for seven days; F: No down-regulation of CD14 expression was observed on histogram of CD14 expressing mononuclear cells after seven days of culture in RPMI (control medium), J82 (control cell line) or WiDR conditioned medium; G: Histogram of CD14 expressing mononuclear cells after seven days of culture in DMEM (control medium) and HT-29 conditioned medium.