Approach to develop a new treatment for patients with implantationfailure using peripheral blood mononuclear cells

在妊娠早期给药ICR小鼠脾细胞可促进假妊娠第2天子宫的接受性。妊娠第4天小鼠的脾细胞比假妊娠第4天和发情第2天小鼠的脾细胞更有效地促进胚胎着床，这表明在供体小鼠中存在胚胎用于脾细胞准备对脾细胞着床的影响很重要。为了探讨脾细胞对子宫内膜分化的影响，我们检测了子宫内LIFmRNA的表达。在去卵巢小鼠中，给予早期妊娠小鼠脾细胞诱导LIFmRNA的表达，雌二醇也观察到这种情况。子宫内膜内注射早期妊娠小鼠脾细胞也能促进胚胎着床。这些发现表明，早期妊娠小鼠脾细胞对着床的影响是独立于卵巢的，并且更直接地促进子宫内膜分化。为了评估免疫细胞是否通过直接作用于胚胎促进胚胎着床，我们采用体外入侵实验研究了早期妊娠妇女外周血单个核细胞（PBMC）对小鼠胚泡BeWo细胞入侵、生长和入侵的影响。PBMC促进了BeWo细胞的侵袭细胞数量，促进了小鼠囊胚的生长和侵袭。此外，PBMC的促进作用来源于非孕妇。这些对BeWo细胞和小鼠囊胚侵袭的促进作用可以通过与r-hCG共培养诱导，提示胚胎存在时观察到的外周免疫细胞的促进作用是由hCG诱导的。基于这些结果，我们提出了一种新的治疗着床失败的方法，即在着床失败的情况下，将经r-hCG预处理的自体外周免疫细胞注入子宫内膜，促进子宫内膜分化和胚胎着床。少

Systemic administration of splenocytes from ICR mice in the early stages of pregnancy promoted the receptivity of the uterus on pseudopregnant day 2. Splenocytes derived from pregnancy day 4 mice more effectively facilitated embryo implantation than splenocytes derived from pseudopregnancy day 4 and diestrous day 2 mice, suggesting that the presence of an embryo in the donor mice for splenocyte preparation is important for the effect of splenocytes on implantation. To investigate the effect of splenocytes on endometrial differentiation, the expression of LIFmRNA in the uterus was examined. In ovariectomized mice, administration of splenocytes from early pregnant mice induced the expression of LIFmRNA which was also observed with estradiol administration. Intra-endometrial injection of splenocytes from early pregnant mice also enhanced embryo implantation. These findings suggest that the effects of splenocytes from early pregnant mice on implantation are independent of the ovary, and di … More rectly facilitate endometrial differentiation. To estimate whether immune cells promote embryo implantation by direct action on embryos or not, we examined the effect of human peripheral blood mononuclear cells (PBMC) derived from early pregnant women on BeWo cell invasion and, outgrowth and invasion of murine blastocysts using in vitro invasion assay. PBMC promoted the invaded cell number of BeWo cells and facilitated outgrowth and invasion of murine blastocysts. Furthermore, the promoting effect of PBMC derived from non-pregnant women. These promotion effects on BeWo cells and murine blastocyst invasion could by induced by the coculture with r-hCG in PBMC derived from non-pregnant women, suggesting that the promoting effect of peripheral immune cells, which are observed in the presence of an embryo, is induced by hCG.Based on these results, we proposed a new treatment for implantation failure in which autologous peripheral immune cells pretreated with r-hCG are injected into the endometrium to facilitate endometrial defferentiation and embryo implantation in cases of implantation failure. Less