小鼠骨髓源性树突细胞和脾源性树突细胞的形态及生物学性能对比

Comparison of mouse bone marrow derived dendritic cells and spleen derived dendritic cellsin morphology and biological functions

目的体外用相同的方法诱导骨髓前体细胞和脾脏细胞生成树突细胞(DCs),从细胞形态和生物学性能两个方面对两者进行比较,为后期小青龙汤干预TSLPDCs体系发挥抗过敏性鼻炎作用筛选合适的DCs细胞。方法从C57小鼠分别获取骨髓细胞和脾细胞,在体外用因子诱导生成DCs,结合流式细胞术(FCM)和酶联免疫吸附测定(ELISA)技术从细胞形态、表面分子的特异性表达、相关因子分泌等几个方面对两者进行比较。结果镜下观察骨髓源性DCs和脾源性DCs均有明显的不规则毛刺状突起,呈集落悬浮状态,但后者集落更明显;扫描电镜下两者均呈薄纱状且有粘连,但后者粘连更大。流式结果显示脾源性DCs纯度为(87.227±4.463)%,骨髓源性DCs纯度为(81.567±2.902)%,两者比较差异有统计学意义(P<0.05)。LPS刺激前脾源性DCs CD11c^+CD83^+表达比例明显低于骨髓源性DCs(P<0.05),LPS刺激后脾源性DCs CD11c^+CD83^+表达比例与骨髓源性DCs比较差异无统计学意义(P>0.05)。ELISA结果显示LPS刺激前脾源性DCs IL-12表达量与骨髓源性DCs IL-12表达量比较差异无统计学意义(P>0.05),LPS刺激后脾源性DCs IL-12表达量与骨髓源性DCs比较差异无统计学意义(P>0.05)。结论骨髓前体细胞和脾细胞在体外经因子诱导后均可获得纯度和性能较好的DCs,且脾源性DCs纯度和性能均高于骨髓源性DCs,但数量明显少于骨髓源性DCs;骨髓源性DCs成熟早于脾源性DCs。

Objective:It is to induce bone marrow progenitor cells and spleen cells to produce dendritic cells(DCs)by the same method in vitro,and compare them from two aspects of cell morphology and biological performance,in order to screen appropriate cells for the latter research of anti allergic rhinitis by the intervention of the Xiaoqinglong Decoction in the TSLP-DCs system.Methods 0x09 Bone marrow cells and spleen cells were obtained from C57 mice and were induced into DCs by GM-CSF and IL-4 in vitro,Flow cytometry(FCM)and enzyme-linked immunosorbent assay(ELISA)were used to compare the cell morphology,specific expression of surface molecules and secretion of related factors in the two aspects.Results 0x09 Under the microscope,the bone marrow-derived DCs and the spleen-derived DCs had obvious irregular burr-like protuberances,and present a state of colonies and suspending,but the latter was more obvious.Under scanning electron microscope,both of them were tulle-like and adhesive,but the latter was more adhesive.Flow cytometry showed that the purity of spleen derived DCs was(87.227±4.463)%,and the purity of bone marrow derived DCs was(81.567±2.902)%,there was significant difference(P<0.05);the expression ratio of CD11c+CD83+in spleen-derived DCs before the stimulation of LPS was significantly lower than that of bone marrow-derived DCs(P<0.05),there was no significant difference in the expression ratio of CD11c+CD83+after the stimulation of LPS between spleen-derived DCs and bone marrow-derived DCs(P>0.05).ELISA results showed that there was no significant difference between the expression of spleen-derived DCs IL-12 and the expression of bone marrow-derived DCs IL-12 before the stimulation of LPS(P>0.05),and there was no significant difference in the expression of IL-12 after the stimulation of LPS between spleen-derived DCs and bone marrow-derived DCs(P>0.05).Conclusion 0x09 DCs with better purity and performance can be obtained in both bone marrow precursor cells and splenocytes after factor induction in vitro,and the purity and performance of spleen derived DCs were higher than that of bone marrow-derived DCs,but the number was significantly less than that of bone marrow-derived DCs;the maturation of bone marrow-derived DCs was earlier than that of spleen-derived DCs.