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目的对粒-单集落刺激因子(GM-CSF)/白介素4(IL-4)或脱氧氟胸腺嘧啶配体(Flt3-L)体外诱导的小鼠骨髓来源树突状细胞(BMDC)亚群和正常小鼠脾脏DC亚群进行比较,探索诱生DC的特性。方法分离Balb/c小鼠骨髓细胞,分别加入含GM-CSF/IL-4或Flt3-L的培养液,体外培养7d,倒置显微镜观察细胞形态,并用流式细胞术检测CD11c、MHCⅡ、CD4、CD8α、CD45RA及Sirp-α分子;利用免疫磁珠从小鼠脾脏细胞中分离DC。结果两组细胞因子均可在体外诱导小鼠骨髓细胞分化发育为未成熟的DC;倒置显微镜下观察可见BMDCs表面有树突状突起,具有典型的DC形态学特点,与Flt3-L诱导的BMDC相比GM-CSF/IL-4诱导的DC体积大,树突长;但是,流式细胞术检测发现Flt3-L体外诱导的BMDCs与小鼠脾脏DC亚群更为相似。结论 GM-CSF/IL-4及Flt3-L均可在体外诱导小鼠骨髓细胞分化发育为DC,且Flt3-LDC亚群与脾脏DC亚群相似,有可能成为体外研究脾脏来源DC的一种方法。
OBJECTIVE: To investigate the effects of granulocyte-monocyte colony stimulating factor (GM-CSF) / interleukin-4 (IL-4) or deoxyfluorene thymidine ligand (Flt3-L) on bone marrow-derived dendritic cells Normal mice spleen DC subpopulations were compared to explore the characteristics of induced DC. Methods Balb / c mouse bone marrow cells were isolated and cultured in vitro. The cells were cultured in vitro for 7 days. The cell morphology was observed under inverted microscope. The expressions of CD11c, MHCⅡ, CD4, CD8α, CD45RA and Sirp-α molecules; immunomagnetic beads were used to isolate DCs from mouse spleen cells. Results Both of the two groups of cytokines could induce the differentiation of mouse bone marrow cells into immature DC in vitro. Under inverted microscope, dendritic protuberances were observed on the surface of BMDCs, showing typical morphological features of DCs. Compared with Flt3-L-induced BMDC The dendrites were longer than DCs induced by GM-CSF / IL-4. However, flow cytometry showed that Flt3-L-induced BMDCs in vitro were more similar to the DC subsets of spleen in mice. Conclusion Both GM-CSF / IL-4 and Flt3-L can induce the differentiation of mouse bone marrow cells into DCs in vitro, and the Flt3-LDC subsets are similar to the spleen DC subsets, which may be one of the methods for studying DCs derived from spleen in vitro method.