Fig. 6
Mixed leukocyte reaction of the differentiated DCs from monocytes. DCs were differentiated from CD14+ monocytes (5 × 105 cells/1.5 mL) in SF-DC Optimal medium, SF-DC Control medium and SF-DC Control + Serum medium for 5 days (Day 0 to Day 5), and then, 20 ng/ml TNF-α or 1 μg/mL lipopolysaccharide (LPS) was added to the corresponding medium to stimulate maturation for 2 days (Day 5 to Day 7). After differentiation, CD209+ DCs were isolated from the differentiated cells using anti-CD209 microbeads on a VarioMACS separator. Then, the isolated CD209+ DCs were cocultured with allogenic CFSE-stained CD3+ T cells at a ratio of 1:2 for mixed leukocyte reactions. After 4 days of coculture, total cells were harvested, and proliferating CD3+ T cells were determined using flow cytometry. A Representative analysis of CFSE expression in cocultured cells. The cells in the dotted boundary represent proliferating CD3+ T cells with decayed CFSE expression. B The percentages of proliferating CD3+ T cells among the total CD3+ T cells. * and *** represent a significant difference of p < 0.05 and p < 0.005, respectively (n = 3). C Growth kinetics of CD3+ T-cell expansion stimulated by coculture with various differentiated DCs (n = 3). The initial CD3+ T-cell density was 1 × 105 cells/mL