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机构地区:[1]首都医科大学基础医学院组织学与胚胎学教研室生殖生物学实验室,北京100069
出 处:《生殖与避孕》2010年第4期223-229,共7页Reproduction and Contraception
基 金:北京市教委科技发展计划项目(项目号:KM200910025005)
摘 要:目的:探讨小鼠子宫自然杀伤细胞(uterine natural killer cells,uNK细胞)与树突状细胞(dendritic cells,DC)之间的相互影响。方法:①妊娠12~13d昆明小鼠子宫腺细胞区组织块培养后,分离出uNK细胞,单独培养(uNK组);②从小鼠骨髓分离得到未成熟DC,单独培养(DC组);③DC和uNK细胞直接接触共培养(共培养组)。观察各组细胞的生长状况,ELISA法检测培养上清液中IL-4、IL-10、IL-12、IL-15及IFN-γ的浓度,流式细胞术检测细胞表面CD40、CD86和CD83的表达。结果:与uNK组和DC组比,共培养组DC和uNK细胞生长聚集现象减少,2种细胞间相互接触,DC细胞表面CD40和CD86表达增高,但是CD83表达仍很低;培养上清液中IL-4、IL-10、IL-12、IL-15及IFN-γ的浓度均增加,尤以IL-4和IL-10增加显著。结论:uNK细胞和DC两者相互作用,DC参与介导uNK细胞的激活,促进其增殖,同时uNK细胞诱导DC呈部分激活状态,使其具有更好的耐受性。Objective: To investigate the influence between uterine natural killer cells (uNK cells) and dendritic cells (DC) of mice in vitro. Methods: 1) uNK cells migrated from blocks of the culturing metrial glands from pregnant mice on day 12-13 and cultured separately (uNK group). 2) Immature DC were originated from bone marrow and cultured separately (DC group). 3) DC and uNK cells co-cultured via direct contact (co-culture group). Cellular morphology was observed. The concentrations of IL-4, IL-10, IL-12, IL-15 and IFN-γ in supernatant were detected by ELISA. The expressions of cell surface markers, CD40, CD86 and CD83, were detected by flow cytometry. Results: Compared with uNK group and DC group, morphology of DC and uNK cells in co-culture group was varied, including the reducing growth mass, and the more cell-cell contacts between DC and uNK cells. The more expressions of CD40 and CD86 were detected, whereas the expressions of CD83 kept at the same low level. The concentrations of IL-4, IL-10, IL-12, IL-15 and IFN-γ in supernatant were increased, the increasing levels of the IL-4 and IL-10 were preponderant (P0.01). Conclusion: DC and uNK cells have interactional effect. DC promote the activations and proliferation of uNK cells. In turn, activated uNKs are able to induce partial activations of DC and make them have better tolerance. Their coordinate may response the benefits to pregnancy at the maternal fetal interface.
关 键 词:子宫自然杀伤细胞(uNK细胞) 树突状细胞(DC) 体外培养
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