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无菌制备BALB/c小鼠骨髓细胞,按常规方法分离纯化出树突状细胞(DCs),采用GM-CSF和IL-4诱导后,以凋亡肿瘤细胞负载未成熟DCs,再分别加入mCD40L-CHO细胞和TNF-α继续培养48h,按常规方法分别制备各发育阶段DCs的超薄切片。用透射电镜观察小鼠DCs在不同发育阶段的超微结构特征,并比较凋亡肿瘤细胞负载的小鼠DCs被CD40L和TNF-α刺激后超微结构的差别。实验结果证实DCs在分化发育成熟中存在异质性;DCs可通过吞噬凋亡的肿瘤细胞负载抗原;CD40配基化对DCs的分化成熟作用优于TNF-α。
BALB / c mouse bone marrow cells were prepared aseptically and dendritic cells (DCs) were isolated and purified by routine methods. After inducing with GM-CSF and IL-4, immature DCs were loaded with apoptotic tumor cells, and mCD40L CHO cells and TNF-α were cultured for 48h. Ultrathin sections of DCs in different developmental stages were prepared by routine methods. The ultrastructural characteristics of mouse DCs at different developmental stages were observed by transmission electron microscopy. The ultrastructural differences of DCs stimulated by CD40L and TNF-α were compared among the DCs loaded with apoptotic tumor cells. The experimental results confirmed that DCs have heterogeneity in differentiation and development. DCs can load antigens through phagocytosis of apoptotic tumor cells. CD40 ligandization is superior to TNF-α in differentiation and maturation of DCs.