小鼠骨髓树突状细胞对胃癌细胞杀伤作用的研究

The Study of Mouse Bone Marrow-derived Dendritic Cells in Vitro Cytotoxicity against Gastric Cancer Cells

目的研究小鼠骨髓树突状细胞的制备及其对胃癌细胞的杀伤作用。方法添加细胞因子培养扩增小鼠骨髓源树突状细胞;流式细胞仪检测DC表面标志物CD11c、CD86的表达;MTT法测定DC对胃癌细胞的杀伤作用。结果培养第7天出现扩增之骨髓源DC,加入TNF-α24 h后,成熟骨髓源DC形成。每只615小鼠平均能分离出3×107个骨髓细胞,一只小鼠的骨髓单个核细胞DC的产量约为7×106个。加入TNF-α48 h后,DC表面标志物CD86阳性表达率较加入TNF-α前增高;CD11c阳性表达率加入TNF–α前后无差别。成熟DC与615小鼠胃癌细胞共孵育24 h、48 h,效靶比分别为10∶1、20∶1、30∶1时,DC对胃癌细胞的杀伤活性随效靶比的增大而增强。结论提取纯化小鼠骨髓细胞,经细胞因子GM-CSF、IL-4、TNF-α的刺激诱导可产生成熟DC,成熟DC对胃癌细胞有较强的杀伤作用,其杀伤活性随效靶比的增大而增强,随共孵育时间的延长杀伤活性有增强的倾向。

Objective To study the preparation of mouse bone marrow-derived dendritic cells in vitro and its Cytotoxicity against gastric cancer cells.Methods Adding cytokines to amplify mouse bone marrow-derived DC and testing the expression of CD11c,CD86 by flow cytometry.MTT colorimetric assay to test the killing effect of DC to gastric cancer cells.Results On the seventh day of culture,amplification of mouse bone marrow-derived DC can be observed.24 hours after adding TNF-α,mature dendritic cells derived from bone marrow can be Seen.An average of 3×107 Ge bone marrow cells can be isolated from every 615 mouses.The bone marrow mononuclear cells of per muose can be induced to generate about 7×106.DCs.After adding TNF-α 48 hours,the positive rates of CD86 after adding TNF-α are higher significantly than that before,the positive rates of CD11c no chang.Mature DCs were cultured with gastric cancer cells for 24 hours and 48 hours respectively.When effector-target ration are 10∶1,20∶1,30∶1,the higher effector-target ration,the more powerful tumocidal killing activities of these DCs to gastric cancer cells.Conclusion By isolateing and amplify bone marrow cells,loading with GM-CSF、IL-4、TNF-α,mature dendritic cells can be obtained.Mature DC have strong killing effect again gastric cancer cells.The killing activities of DC to gastric cancer cells increase with effector-target ration.The tendency of increase can be observed with incubation times extended.