There is a constant need for improving embryo culture conditions in assisted reproduction. One possibility is to use mesenchymal stem/stromal cells derived from menstrual blood (mbMSCs), with an endometrial origin. In this study, we sought to analyze the expansion of mouse embryos in a direct coculture model with mbMSCs. Our results showed that after five passages, mbMSCs presented a spindle-shaped morphology, with surface markers that were comparable with the normal mesenchymal cell phenotype. mbMSCs could differentiate into adipogenic and osteogenic lineages and secrete angiopoetin-2 and hepatocyte growth factor. The coculture experiments employed 103 two-cell-stage embryos that were randomly divided into two groups: control (n = 50), embryos cultured in GV-Blast medium, and cocultured mbMSCs (n = 53), embryos cocultured with GV-Blast and mbMSCs. Typically, two to three embryos were placed in a well with 200 μL of culture medium and observed until developmental day 5. After 5 days, the cocultured group had more embryos in the blastocyst stage (69.8%) when compared with the control group (30%) (p < 0.001). It was also found that nearly 57% of blastocysts in the cocultured group reached the hatching stage, while only 13% achieved this stage in the control group (p < 0.001). Analyses of cultured mbMSCs and growth media, in the presence or absence of an embryo, were also performed. Immunofluorescence detected similar levels of collagen I and III and fibronectin in both mbMSCs and cocultured mbMSCs, and similar amounts of growth factors, VEGF, PDGF-AA, and PDGF-BB, were also observed in the conditioned medium, regardless of embryo presence. The present study describes, for the first time, an easy, noninvasive, and autologous method that could potentially increase blastocyst growth rates during assisted reproductive procedures (i.e., in vitro fertilization). It is proposed that this mbMSC coculture strategy enriches the embryonic microenvironment and promotes embryo development. This technique may complement or replace existing assisted reproduction methods and is directly relevant to the field of personalized medicine.

中文翻译：

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人经血来源的间充质细胞改善小鼠胚胎发育。

一直需要在辅助生殖中改善胚胎培养条件。一种可能性是使用源自月经血（mbMSCs）的具有子宫内膜来源的间充质干/基质细胞。在这项研究中，我们试图分析与mbMSCs直接共培养模型中小鼠胚胎的扩增。我们的结果表明，经过五次传代，mbMSCs呈纺锤形形态，其表面标志物可与正常的间充质细胞表型相媲美。mbMSCs可以分化为成脂和成骨谱系，并分泌血管生成素2和肝细胞生长因子。共培养实验采用103个两细胞阶段的胚胎，将其随机分为两组：对照（n  = 50），在GV-Blast培养基中培养的胚胎和mbMSCs（n = 53），将胚胎与GV-Blast和mbMSCs共培养。通常，将两到三个胚胎放在装有200μL培养基的孔中，并观察到发育第5天。5天后，与对照组（30只）相比，共培养组在胚泡期的胚胎更多（69.8％）。 ％）（p  <0.001）。还发现在共培养组中接近57％的囊胚达到孵化阶段，而在对照组中只有13％达到了该阶段（p  <0.001）。在有或没有胚胎的情况下，还对培养的mbMSCs和生长培养基进行了分析。免疫荧光法检测到mbMSCs和共培养的mbMSCs中的胶原蛋白I和III和纤连蛋白的水平相近，并且生长因子VEGF，在条件培养基中也观察到PDGF-AA和PDGF-BB，无论是否存在胚胎。本研究首次描述了一种简单，无创且自体的方法，该方法可能在辅助生殖程序（即体外受精）过程中提高胚泡的生长速度。有人提出这种mbMSC共培养策略可以丰富胚胎微环境并促进胚胎发育。该技术可以补充或替代现有的辅助生殖方法，并且与个性化医学领域直接相关。