体外诱导成熟树突状细胞的研究

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％、67.2％、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能表达高高水平的B7－1和CD40分子，这为大量获得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，为开展以DC为基础的各种免疫治疗研究奠了良好的基础。

Research of maturedendritic cells induced in vitro

Objective: To study the suitable condition of inducing maturedendritic ce lls (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 TN F-α affecting, the mBmDC, fLDC and fetal bone marrow DC (fBm DC) grow as typic a l DC which cytoplasm is protrusion like polythoorn. The 5d rates of harvest fro m 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 wit h 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, f BmDC 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.