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目的:建立小鼠子宫淋巴细胞分离方法,并初步观察小鼠子宫uNK细胞的特性。方法:分别用机械法和酶消化法制备单细胞悬液分离小鼠子宫淋巴细胞,用二色或三色荧光标记技术检测子宫uNK细胞的比例及相关特征。结果:机械法分离淋巴细胞技术相对稳定、纯度较高;酶消化法未降低DX5 +细胞所占的比例,但酶的消化过程容易使淋巴细胞活化水平升高,表现为活化分子CD6 9的表达上调。子宫中存在大量的uNK细胞(NK1 1+CD3-或DX5 +CD3-细胞) ,小鼠怀孕早期和中期,uNK细胞占子宫淋巴细胞的比例随着怀孕天数的增长而增长,到怀孕的第10天达到高峰(uNK细胞比例达到4 8 .74 % ) ,之后比例开始迅速下降。结论:成功建立了小鼠子宫uNK细胞分离方法,子宫uNK细胞作为重要的天然免疫淋巴细胞,可能参与妊娠子宫对半同种异体胚胎的保护性免疫反应。
Objective: To establish a mouse uterine lymphocyte separation method and to observe the characteristics of mouse uterus uNK cells. Methods: Mouse uterine lymphocytes were isolated by mechanical and enzymatic digestion method respectively. The proportion of uNK cells and their related features were detected by two-color or three-color fluorescent labeling. RESULTS: The technique of mechanical separation of lymphocytes was relatively stable with high purity. Enzymatic digestion did not reduce the proportion of DX5 + cells, but the enzymatic digestion facilitated the activation of lymphocytes, which showed the expression of CD6 9 Increase. In the uterus, there are a large number of uNK cells (NK1 1 + CD3- or DX5 + CD3- cells). The proportion of uNK cells in the uterus lymphocytes increased with the number of pregnant days in the early and middle stages of pregnancy to 10 The highest peak day (uNK cell ratio reached 48.74%), after which the ratio began to decline rapidly. Conclusion: The uNK cell isolation method has been established successfully. Uterine uNK cells are important innate immune lymphocytes and may participate in the protective immune response to the semi-allogeneic embryos of pregnant uterus.