One of the effective treatments in women with recurrent implantation failure (RIF) is the use of immune cells to facilitate embryo implantation. Previous studies have shown that intrauterine transmission of peripheral blood mononuclear cells (PBMC) increased the embryo implantation rate. In this study using B6D2F1 (C57BL/6 × DBA2) mice, a fragment of the granulocyte macrophage colony-stimulating factor (Gm-csf) gene was cloned into an enhanced green fluorescent protein vector (pEGFP-N1) and then transfected into PBMC. The protein level of GM-CSF was evaluated in the transfected PBMC and untransfected PBMC by ELISA. Attachment of mouse embryos and the mRNA expression levels of leukemia inhibitory factor (Lif), vascular endothelial growth factor (Vegf), matrix metalloproteinase 9 (Mmp9), Gmcsf-receptor (Gmcsf-r) and interleukin 6 (Il6) in vitro were assessed by real-time PCR in endometrial cells. To determine the pregnancy rate and number of implantation sites in vivo, the mouse uterine horns were analyzed on Day 7.5 post coitum. A greater amount of GM-CSF was produced in PBMC transfected with recombinant vector (552 pg/mL) compared with the untransfected PBMC (57 pg/mL) and PBMC transfected with empty vector (34 pg/mL) (P < 0.05). The data showed that the embryo attachment rate and mRNA expression levels (Vegf [1.7-fold], Mmp9 [1.4-fold], Lif [1.5-fold], Gm-csf r [1.6-fold] and Il6 [1.2-fold]) in the in vitro study (P < 0.01), pregnancy rate (P < 0.01) and number of implantation sites (P < 0.01) in the in vivo investigation (P < 0.05) were increased in PBMC transfected with recombinant vector compared with the PBMC group. The study demonstrated that, in mice, endometrium immunotherapy with transfected PBMC that contained recombinant GM-CSF before embryo implantation was effective in improving embryo implantation and endometrial receptivity.

中文翻译：

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GM-CSF宫内给药转染的外周血单个核细胞对小鼠胚胎着床和妊娠率的作用。

复发性植入失败（RIF）妇女的有效治疗方法之一是使用免疫细胞促进胚胎植入。先前的研究表明，外周血单个核细胞（PBMC）在子宫内的传播会增加胚胎的植入率。在这项使用B6D2F1（C57BL / 6×DBA2）小鼠的研究中，将粒细胞巨噬细胞集落刺激因子（Gm-csf）基因的片段克隆到增强的绿色荧光蛋白载体（pEGFP-N1）中，然后转染到PBMC中。通过ELISA评估转染的PBMC和未转染的PBMC中GM-CSF的蛋白水平。小鼠胚胎的附着和白血病抑制因子（Lif），血管内皮生长因子（Vegf），基质金属蛋白酶9（Mmp9）的mRNA表达水平，通过实时PCR在子宫内膜细胞中评估了体外Gmcsf受体（Gmcsf-r）和白介素6（Il6）。为了确定体内的怀孕率和植入部位的数量，在宫腔放置后第7.5天对小鼠子宫角进行了分析。与未转染的PBMC（57 pg / mL）和空载体（34 pg / mL）转染的PBMC相比，重组载体（552 pg / mL）转染的PBMC中产生的GM-CSF量更大（P <0.05）。数据显示胚胎附着率和mRNA表达水平（Vegf [1.7倍]，Mmp9 [1.4倍]，Lif [1.5倍]，Gm-csf r [1.6倍]和Il6 [1.2倍] ）的体外研究（P <0.01），体内研究的妊娠率（P <0.01）和植入部位数目（P <0.01）（P <0。与PBMC组相比，用重组载体转染的PBMC中05）增加。该研究表明，在小鼠体内，在胚胎植入前用转染的PBMC进行子宫内膜免疫治疗是有效的，该PBMC含有重组GM-CSF，可以改善胚胎植入和子宫内膜的接受性。