Differentiation of dendritic cells from embryonic stem cells

AIM：To investigate the method of directed differentiation dendritic cells (DC) from embryonic stem cells,and to amplify high purity dendritic cells in vitro for immunolgical therapy.METHODS：E14 embryonic stem cell line was generated ES-DC in complete medium supplemented with GM-CSF and IL-3.Flow cytometry was used to determine CD11c,CD80,CD86,MHC-II cell-surface phenotype in immatured ES-DC.Lipopolysaccharide (LPS) was added to induce the ES-DC maturation.The matured ES-DC was harvested 24 hours later to identifying with morphology and transmission electron microscopy.The phenotype of matured ES-DC was analyzed by flow cytometry and compared with the immatured ES-DC.The antigen presenting was evaluated by mixed lymphocyte responses (MLR).RESULTS：The ES-DC had obviously dendritic processes under scanning electron microscope.The immature DCs expressed low level of CD11c (4.33±0.23)%,CD80 (7.62±0.19) %,CD86 (4.77±1.22) % and MHC-II (9.68±0.15) %.The mature DCs expressed higher level of CD11c (47.36±2.68)%,CD80 (74.40±1.47) %,CD86 (29.77±2.00) % and MHC-II (87.56±2.75) %.MLR showed that ES-DCs effectively stimulated lymphocyte proliferation.CONCLUSIONS：These results provide evidence that dendritic cells can be generated from E14 embryonic cells with the stimulations of GM-CSF and IL-3.The differentiated cells expresse high level of CD11c,CD80,CD86,MHC-II and effectively stimulate lymphocytes to proliferate.ES cells may become new origin of DC for immunotherapy.