两种来源树突状细胞生物学特性的比较

Comparison of biologic characteristics between dendritic cells from two sources

目的体外诱导小鼠胚胎干细胞(ESC)以及骨髓(BM)单个核细胞分化为树突状细胞(DC),并对其生物学特性进行比较。方法悬浮培养诱导小鼠ESC生成拟胚体(embryonic bodies,EB),取14d生长良好的EB转移至组织培养皿中,添加粒细胞-巨噬细胞集落刺激因子(GM-CSF)以及IL-3,培养2周后收获DC。同时联合应用GM-CSF、IL-4诱导培养小鼠骨髓单个核细胞分化为DC。并从形态学、表型及功能方面对两者进行检测。结果ESC-DC、BM-DC经磷酸脂多糖(LPS)刺激成熟后其表面均显示出明显的树枝状突起,高表达CD11c、CD86及MHC-Ⅱ类分子,并具有极强的刺激同种异基因淋巴细胞增殖的能力。结论ESC-DC具有与BM-DC相同的生物学特性,为DC的功能研究以及进一步制备相应的白血病疫苗提供又一可靠来源。

Objective To induce dendritic cells from mice embryonic stem cells （ESCs） and bone marrow mononuclear ceils in vitro, and compare their biologic characteristics. Methods Embryonic bodies （EBs） were induced from mice ESCs suspending culture in petri dishes. After cultured for 14 days, EBs were transferred onto tissue culture flasks, and GM-CSF and IL-3 （each at 25 ng/ml） were added. Immature dendritic cells （DCs） were taken for study after cultured for 2 weeks. Mononuclear cells isolated from mice bone marrow were induced into dendritic cells by culturing in GM-CSF and IL-4 （each at 25 ng/ml）. Morphology, phenotype and functions of the two differently derived dendrite cells were examined. Results ESC-DCs and BM-DCs were induced to mature through exposure to lipopolysaccharide （LPS） and exhibited extensive dendrites on their surfaces. Both types of the DCs showed high expression of CD11c, MHC-Ⅱ , CD86 and strong capacity to stimulate primary Tcell responses in mixed leukocyte reaction （MLR）. Conclusions ESC-DC shared the same biologic characteristics with BM-DC, offering a new reliable source for research on the function of DC and further provides a resource for leukemic vaccine.