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Objective: To compare C57BL / 6 of mice marrow-derived dendritic cells (dendritic cell, DC) and genetically modified DC2.4 morphology and function. And to study the the different doses Dengzhanhua extract DC2.4 mature state and function. Methods: C57BL / 6 mice long bone in the bone marrow cells, semi-adherent method DC precursors, granulocyte - macrophage colony-stimulating factor (GM-CSF) and interleukin-4 ( culture, IL-4) synergistically induced stimulation with lipopolysaccharide (LPS) in DC2.4 and the first 7d DC 48h, cell morphology was observed and the detection of the IL-12 concentration and cell surface markers the CDl 1c, CD80, CD86 and MHC-Ⅱ; will develop the DC2.4 divided into negative group (simple culture medium), positive group (LPS induced maturation) and experimental group (in addition to LPS was added, and then added Dengzhanhua extract). The experimental group was divided into 1 to 4 four groups were added to different doses (50, 100, 150 and 200μg/ml) Dengzhanhua extract of cultured 3d observe the morphological changes detected CDl 1c, CD40, MHC-Ⅱ, IL -12 mixed lymphocyte culture experiments to stimulate T cell proliferation. 3d visible results: mouse marrow-derived DC is DC2.4 compare: inverted microscope DC2.4 DC morphology cultured to adherent mononuclear macrophages and semi-adherent cells, became larger, irregularly shaped was the star, tadpole-shaped or spindle, a large number of projections; 4 ~ 7d visible part of the cells increased off the wall, was characteristic star-shaped projections ranging from 2 to 5, some longer, but still see fusiform; the first 7d adding LPS 48 h, DC a lot of off the wall, and the branch is increased, more coarse. DC2.4 was semi-adherent state, the shape of a star, tadpole-shaped or spindle-shaped, similar to the mouse marrow-derived dendritic cell morphology. With LPS, increased dendritic branching, thickening. Concentration of DC IL-12 after LPS stimulation (32.01 ± 2.62) pg / ml, and the difference was statistically significant (P <0.01 and training to 9d mouse bone marrow-derived DC group (14.36 ± 1.98) pg / ml) ; DC2.4 IL-12 after LPS stimulation concentration (29.92 ± 5.76) pg / ml higher than the of DC2.4 group (14.98 ± 4.75) pg / ml (P <0.05). FACS analysis, DC2.4 cells express the CDl 1c, CD80, CD86 and MHC-Ⅱ molecules, respectively (1.62 ± 0.51)% and (2.03 ± 0.15)% and (8.19 ± 1.13)%, (13.88 ± 12.62)% ; after LPS-induced DC2.4 group (5.54 ± 1.08)%, (2.42 ± 0.20)%, (9.59 ± 1.27)%, (31.45 ± 12.09)%, the CDl 1c and the MHC-Ⅱ difference was statistically significant. (P <0.05). Cultured until 48 h after the first-9d mouse marrow derived DC and LPS stimulation, DC express CDl 1c, CD80, CD86 and MHC-Ⅱ molecules in cells, respectively (55.11 ± 4.48)%, (40.68 ± 3.77)%, ( 29.91 ± 3.59)%, (41.27 ± 2.95)% and (84.25 ± 4.62)%, (74.61 ± 4.94)%, (61.50 ± 5.10)%, (57.41 ± 3.31)%, were statistically significant (P <0.01 ). See an inverted microscope DC2.4 DC2.4 by erigeron culture extracts treated was semi-adherent, star, tadpole-shaped or fusiform, DC morphology. With LPS, the increase in dendritic branching, thickening, nearly circular, polygonal or characteristic star grows louder, membrane cytoplasm dendritic protrusions formed. Cell extract under relatively naive, sparse, and the difference is more obvious in high doses. TEM observations DC2.4 surface processes more visible, the nucleus and prominent nucleoli lot of phagocytic vesicles within the cytoplasm, also visible homogeneous lipid droplets in the cytoplasm. Multi-branch, scanning electron microscope after LPS stimulation surface protrusions. IL-12 levels were measured in the positive group measured the level of IL-12 the 29.92 ± 5.76pg/ml, compared with the negative group and the experimental group, the difference was statistically significant (P <0.01). In addition to the comparison between the lowest and highest dose group P <0.05, significant difference, the remaining groups, to compare P> 0.05, no significant difference. The DC2.4 markers tested Dengzhanhua extract objects in each dose group, in addition to the experimental group 2 and 3 no significant difference in the CD11c expression from high to low, and groups have a significant difference (P <0.05 P <0.01), CD40 in the experimental group, group 1 is higher than the other groups (P <0.01), the group 2,3,4 no significant difference. MHC-Ⅱ in the experimental group 1 than group 4 (P <0.05), I experimental group had no significant difference (P> 0.05). Mixed lymphocyte culture experiments in addition to the experimental group 3 and 4 was no significant difference in the 10:1 ratio, stimulation index with dose lower (P <0.01) increase from high to low; ratio of 50:1 and 100:1 the group 2,3,4 no difference between group 1 than in each group (P <0.01). Conclusion: 1.DC2.4 C57BL / 6 most of the features of marrow-derived dendritic cells as an in vitro alternative model. And high purity, save time, training costs low. 2.LPS having a role in promoting primary cells and DC2.4 mature. In cell morphology, surface markers, and the secretion of cytokines, the stimulatory effects of the DC and DC2.4 not entirely consistent. 3.DC2.4 mature state and function of cell morphology, expression of surface markers and cytokines are closely related. 4 the. Dengzhanhua extract can inhibit DC2.4 mature to increase with the dose of the extract, the inhibition gradually increased. 5.DC2.4 features and mature state are closely related, with DC2.4 the mature, antigen presentation of the cells gradually strengthened.
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