人自体单个核细胞对体外胚胎和子宫内膜共培养模型中白血病抑制因子及其受体表达的影响  

Effects of human autologous mononuclear cells on expression of leukemia inhibitory factor and its receptor in co-culture model of embryo and endometrium in vitro

作　　者：禹果[1] 项云改[1] 李艳[1] 楚喜英 仝丽丽[1] 窦倩[1] 卢娜[1] 谭丽[1] Yu Guo;Xiang Yungai;Li Yan;Chu Xiying;Tong Lili;Dou Qian;Lu Na;Tan Li(Department of Reproductive Medicine,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China)

机构地区：[1]郑州大学第二附属医院生殖医学部,450014

出　　处：《中华生殖与避孕杂志》2020年第2期124-128,共5页Chinese Journal of Reproduction and Contraception

基　　金：河南省科技厅科技攻关项目(162102310207);河南省卫计委普通攻关项目(201702085);河南省科技厅基础研究(142300410240)。

摘　　要：目的建立胚胎和子宫内膜共培养模型,探讨人自体单个核细胞(PBMCs)对培养液中白血病抑制因子(LIF)和胚胎表面LIF受体(LIFR)表达的影响。方法于2015年5月至2016年5月期间,选择在郑州大学第二附属医院行辅助生殖技术助孕反复种植失败患者的内膜12例,分离出子宫内膜细胞。收集其外周血分离出PBMCs。收集2009年至2014年行辅助生殖技术助孕成功且分娩的患者的废弃胚胎。研究分为实验组(内膜-胚胎-PBMCs共培养)和对照组(内膜-胚胎共培养)。共培养系统中加入含雌孕激素培养液当日为培养第0日。应用酶联免疫法检测培养后第1、第3、第5日两组培养液中LIF的浓度;应用免疫荧光方法检测实验组和对照组培养第5、第6日所形成囊胚表面LIF受体的表达情况,用Image J软件分析免疫荧光图像。结果培养第3日实验组和对照组培养液中LIF浓度差异无统计学意义(P>0.05),培养第5日时,实验组培养液中LIF浓度[(2840.02±240.51)ng/L]明显高于对照组[(2411.35±311.63)ng/L](P=0.001)。囊胚LIFR的免疫荧光吸光度(A)值实验组大于对照组(0.2558±0.0372比0.1906±0.0404,P=0.001)。结论在体外子宫内膜-胚胎共培养模型中,人自体PBMCs能提高子宫内膜LIF的表达,同时促进囊胚表面LIFR受体的形成,在胚胎着床过程中起重要作用。Objective To investigate the effects of human autologous mononuclear cells(PBMCs)on the expression of leukemia inhibitory factor(LIF)and leukemia inhibitory factor receptor(LIFR)on embryonic surface in co-culture model of embryo and endometrium.Methods From May 2015 to May 2016,endometrial glandular epithelial cells were isolated from 12 patients diagnosed as repeated implantation failure in the Second Affiliated Hospital of Zhengzhou University.PBMCs cells were isolated from their peripheral blood.We collected abandoned embryos(signed informed consent of abandoned embryos)from patients who had successfully delivered by assisted reproductive technology from 2009 to 2014.The study was divided into two groups.The experimental group was endometrium-embryo-PBMCs co-culture,and control group was endometrium-embryo co-culture.The day on which the culture medium containing estrogen and progesterone was added was regarded as the 0th day of culture.The concentration of LIF was detected by enzyme linked immunosorbent assay on the 1st,3rd and 5th day after culture.The expression of LIFR on blastocyst surface was detected by immunofluorescence on the 5th and 6th day after culture in experimental group and control group.Immunofluorescence images were analyzed by Image J software.Results On the 3rd day of culture,there was no significant difference in LIF concentration between experimental group and control group(P>0.05).On the 5th day of culture,LIF concentration in experimental group[(2840.02±240.51)ng/L]was significantly higher than that in control group[(2411.35±311.63)ng/L](P=0.001).The immunofluorescence aborsance(A)value of LIFR in blastocysts of experimental group was higher than that of control group(0.2558±0.0372 vs.0.1906±0.0404,P=0.001).Conclusion Human PBMCs can enhance the expression of LIF in endometrium and promote the expression of LIFR on blastocyst surface,which plays an important role in embryo implantation.

关 键 词：人外周血单个核细胞 反复种植失败 白血病抑制因子 共培养 

分 类 号：R714[医药卫生—妇产科学]