【摘要】 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。更多还原

【Abstract】 Objective:To isolate the uterine endometrium lymphocytes and study the phenotypic characters of murine uNK cells.Methods:Mechanical dissection and digestive enzymes were used to prepare cell suspension respectively,and the proportion and phenotypic characters of murine uNK cells were analyzed with two or three colors immunofluorescent-labeling.Results:Compared to digestive enzymes,the mechanical dissection yielded a more purer population of uNK cells.Digestive enzymes did not decrease the proportion of the DX5+ cells in the uterine endometrium,but the digestive enzymes increases the expression levels of CD69 of uNK cells,the results of mechanical dissection is stable.We found that uterine NK cells are the most abundant lymphocytes in murine implantation sites.In the early and middle gestation,NK cells became initially moarked after implantation(gd=4-6),peaked(43.065±12.93298%) at gd10,and quickly decreased after gd12.Conclusion:Uterine nataural killer cells are successfully purified,uNK cells may be involved in the mechanisms of protecting the hemiallogeneic fetal cells from the aggressive response.更多还原