体外诱导成熟树突状细胞的研究

Research of mature dendritic cells induced in vitro

目的 :探讨在体外从干细胞中诱导出成熟DC的适宜环境。方法 :分别将人胎肝、骨髓和脾细胞以及小鼠骨髓和脾细胞在体外用GM CSF和TNF α诱导 ,观察了第 3、5、7天DC的收获情况。进一步用S P免疫细胞化学染色检测了mBmDC和fLDC有关分子表达。结果 :在体外通过GM CSF和TNF α的作用 ,小鼠骨髓、人胎肝、骨髓细胞呈典型的毛刺状胞浆突起 ,第5天的收获率分别为 :39 5 %、6 7 2 %、12 9% ,而基本不能从脾细胞中诱导出成熟DC ,获得的DC能与MAbCD80、MAbCD40呈强阳性反应。结论 :在GM CSF和TNF α的共同作用下 ,能在体外从人胎肝、骨髓和小鼠骨髓干细胞中获得成熟DC ,其DC能表达高水平的B7 1和CD40分子 ,这为大量获取DC提供了一种简便可行的手段 ,为研究DC的生物学特征及其抗原提呈功能提供了丰富的材料 。

Objective:To study the suitable condition of inducing mature dendritic cells (DC) from stem cells in vitro Methods:With GM CSF and TNF α, the cells of liver, bone marrow, spleen from human fetals and bone marrow, spleen from mice were cultured The rates of harvest DC were counted on 3 d, 5 d and 7 d of culture Some molecular expression of the mice bone marrow DC (mBmDC) and fetal liver DC (fLDC) were detected by S P immunocytochemical stain Results:By GM CSF and TNF α affecting, the mBmDC, fLDC and fetal bone marrow DC (fBm DC) grow as typical DC which cytoplasm is protrusion like polythoorn The 5d rates of harvest from mBmDC, fLDC and fBmDC were 39 5% 67 2% and 12 9%, but there were few mature DCs induced from spleen cells The mBmDC, fLDC and fBmDC were strong positive with MAb CD80 and MAb CD40 Conclusion:Mature DCs can be induced in vitro from the stem cells of fetal liver, bone marrow and mice bone marrow with cooperate with GM CSF and TNF α The high lever of B7 1 and CD40 were expressed in fLDC, fBmDC and mBmDC These results have provide a available and simply method of obtaining a lot of DCs It is settled a good basis for DC immunotherapy with increasing cellular immunity