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目的探索不同级别小鼠来源的骨髓细胞对体外制备优势DC2细胞的影响。方法取SPF级和健康普通级C57BL/6小鼠股、胫骨骨髓细胞及脾细胞,设细胞因子诱导的实验组、低浓度GM-CSF对照组、无外源细胞因子对照组及脾细胞组。骨髓细胞采用GM-CSF,IL-4,Flt3L和SCF的不同细胞因子浓度和组合体外培养诱导。脾细胞培养2d去除大部分淋巴细胞,第3天采用流式细胞术4色荧光标记法分析细胞表型,骨髓细胞诱导的各组同样于第3天采用流式细胞术分析细胞表型,比较不同级别小鼠骨髓细胞所诱导的DC2细胞的表型和比例。结果SPF来源的小鼠骨髓细胞诱导的各组第3d在CD11c+的DC细胞群中CD8α+DC2所占比例及不成熟DC2的比例均高于普通级小鼠,显示普通鼠来源的骨髓细胞更倾向于产生成熟的DC,不易诱导产生高比例的不成熟的DC2。对两种动物脾细胞DC2的分析结果表明,普通鼠脾脏中CD8α-MHCⅡ+的成熟DC1比例为75.58%,远远超过SPF鼠的30.60%。结论小鼠的微生物级别可影响耐受性DC的诱导效果,采用SPF级小鼠来源的骨髓细胞体外制备优势DC2细胞效果优于普通级鼠,原因可能与普通鼠接触微生物较多,免疫系统更易于产生免疫应答而非免疫耐受有关。
Objective To explore the effect of different levels of bone marrow cells derived from mice on the preparation of DC2 cells in vitro. Methods C57BL / 6 mice, tibial bone marrow cells and splenocytes of SPF-class and healthy normal-grade mice were divided into three groups: experimental group induced by cytokines, control group with low concentration of GM-CSF, control group without exogenous cytokines and spleen cells. Bone marrow cells were induced using different concentrations of cytokines GM-CSF, IL-4, Flt3L and SCF in vitro and in vitro. Spleen cells were cultured for 2 days to remove most of the lymphocytes. On the 3rd day, the cell phenotypes were analyzed by flow cytometry 4-color fluorescent labeling. The cells induced by bone marrow cells were also analyzed by flow cytometry on the 3rd day. The Phenotypes and Proportions of DC2 Cells Induced by Different Levels of Mouse Marrow Cells. Results The percentage of CD8α + DC2 and the proportion of immature DC2 in DCs derived from CD11c + cells induced by SPF-derived mouse bone marrow cells were higher than those in normal mice on day 3, indicating that the common mouse-derived bone marrow cells tended to be more To produce mature DC, it is not easy to induce a high proportion of immature DC2. The results of DC2 analysis showed that the percentage of mature DC1 in CD8α-MHCⅡ + was 75.58% in the spleens of normal mice, far exceeding 30.60% of SPF mice. CONCLUSION: The microbiological level of mice can affect the inducing effect of tolerogenic DCs. The advantage of producing superior DC2 cells in vitro by SPF-derived mouse bone marrow cells is better than that of common rats. The reason may be that the immune system is more Easy to produce immune response rather than immune tolerance related.