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[目的]探讨建立一种简易、高效的体外分离培养、纯化大鼠骨髓来源树突状细胞(DC)的方法.[方法]采用粒细胞-巨噬细胞集落刺激因子(GM-CSF)+白细胞介素-4(IL-4)分离诱导大鼠骨髓来源造血前体细胞7d后得未成熟树突状细胞(immature dendritic cell,imDC),流式细胞仪检测免疫磁珠纯化前后imDC表面OX62阳性表达率.然后脂多糖(LPS)作用2d刺激imDC成熟得成熟树突状细胞(mature dendritic cell,mDC),流式细胞仪检测imDC及mDC细胞表面CD86、CD80、RT1B阳性表达率,ELISA法检测两组细胞的IL-12分泌水平,体外混合淋巴细胞反应(mixed lymphocyte reaction,MLR)测定两组细胞促淋巴细胞增殖的能力.[结果]免疫磁珠纯化后imDC表面OX62阳性表达率为91.9％,明显高于纯化前的82.8％(P＜0.05);imDC表面CD86、CD80、RT1B阳性表达率分别是11.48％、6.25％、77.2％,经LPS刺激成熟后的mDC表面CD86、CD80和RT1B阳性表达率分别为91.38％、93.54％、98.6％;imDC细胞的IL-12分泌水平为(36±9)pg/mL,明显低于mDC(228±27)pg/mL(P＜0.01);MLR结果中mDC组刺激T淋巴细胞增殖的能力明显高于imDC组.[结论]本实验通过细胞形态观察、表面分子标志和功能状态三方面相结合对所培养细胞进行鉴定,证实联合应用GM-CSF+ IL-4可体外诱导出高纯度、数量多的DC;通过免疫磁珠纯化可获得相对数量多、纯度高的DC以满足实验需要.“,”[Objective] To establish a simple and efficient in vitro isolation and culture system for bone marrow-derived precursor cells (DC) in rats.[Methods] Granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) were used to induce and separate rat bone marrow hematopoietic progenitor cells to get immature dendritic cells after 7 d (imDC),immunomagnetic flow cytometry was used before and after purification of the surface of imDC to test positive expression rate of OX62.Lipopolysaccharide (LPS) lasted for 2 D to stimulate imDC maturation to get mature dendritic cells (mature dendritic cell,mDC),flow cytometry was used to detect imDC and mDC cell surface CD86,CD80,RT1B positive expression rate;ELISA was used to detect the secretion of IL-12 in vitro in two groups of cells,mixed lymphocyte reaction (MLR) was used to determine lymphocyte proliferation ability of cells in two groups.[Results] Purified imDC surface OX62 positive expression rate was 91.9％,significantly higher than that before purification 82.8％ (P ＜0.05);imDC,CD80,CD86 on the surface of the positive expression rates of RT1B were 11.48％,6.25％,77.2％,stimulated by LPS after the maturity of the mDC surface CD86,CD80 and RT1B positive expression rates were 91.38％,93.54％,98.6％ imDC;cell IL-12 secretion level (36±9) pg/mL,was significantly lower than that of mDC (228 ±27) pg/mL (P ＜0.01) MLR results in mDC group;ability to stimulate T lymphocyte proliferation was significantly higher than that of the imDC group.[Conclusion]In this experiment,the cultured cells were identified by three aspects (cell morphology observation,surface molecular markers and functional status),and it was proved that the combination of GM-CSF+ IL-4 could induce high purity and large numbers of DC in vitro;through the purification of immunomagnetic beads,a relatively large number of DC with high purity can be obtained to meet the experimental requirements.