Thin endometrium adversely affects reproductive success rates with fertility treatment. Autologous transplantation of exogenously prepared endometrium can be a promising therapeutic option for thin endometrium; however, endometrial epithelial cells have limited expansion potential, which needs to be overcome in order to make regenerative medicine a therapeutic strategy for refractory thin endometrium. Here, we aimed to perform long-term culture of endometrial epithelial cells in vitro. We prepared primary human endometrial epithelial cells and endometrial stromal cells and investigated whether endometrial stromal cells and human embryonic stem cell-derived feeder cells could support proliferation of endometrial epithelial cells. We also investigated whether three-dimensional culture can be achieved using thawed endometrial epithelial cells and endometrial stromal cells. Co-cultivation with the feeder cells dramatically increased the proliferation rate of the endometrial epithelial cells. We serially passaged the endometrial epithelial cells on mouse embryonic fibroblasts up to passage 6 for 4 months. Among the human-derived feeder cells, endometrial stromal cells exhibited the best feeder activity for proliferation of the endometrial epithelial cells. We continued to propagate the endometrial epithelial cells on endometrial stromal cells up to passage 5 for 81 days. Furthermore, endometrial epithelium and stroma, after the freeze-thaw procedure and sequential culture, were able to establish an endometrial three-dimensional model. We herein established a model of in vitro cultured endometrium as a potential therapeutic option for refractory thin endometrium. The three-dimensional culture model with endometrial epithelial and stromal cell orchestration via cytokines, membrane-bound molecules, extracellular matrices, and gap junction will provide a new framework for exploring the mechanisms underlying the phenomenon of implantation. Additionally, modified embryo culture, so-called “in vitro implantation”, will be possible therapeutic approaches in fertility treatment.

中文翻译：

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子宫内膜上皮的子宫内膜再生：子宫内膜基质作为饲养的同源编排

子宫内膜变薄对生育治疗的生殖成功率产生不利影响。自体准备的内膜子宫内膜移植可能是子宫内膜薄的一种有希望的治疗选择。然而，子宫内膜上皮细胞具有有限的扩增潜力，为了使再生医学成为难治性薄内膜的治疗策略，必须克服这一潜力。在这里，我们旨在进行体外子宫内膜上皮细胞的长期培养。我们准备了原代人子宫内膜上皮细胞和子宫内膜基质细胞，并研究了子宫内膜基质细胞和人胚胎干细胞衍生的饲养细胞是否可以支持子宫内膜上皮细胞的增殖。我们还研究了使用融化的子宫内膜上皮细胞和子宫内膜基质细胞是否可以实现三维培养。与饲养细胞的共培养显着提高了子宫内膜上皮细胞的增殖速率。我们将小鼠胚胎成纤维细胞上的子宫内膜上皮细胞连续传代，直到第6代传代4个月。在人类来源的饲养细胞中，子宫内膜基质细胞对子宫内膜上皮细胞的增殖表现出最佳的饲养活性。我们继续在子宫内膜基质细胞上繁殖子宫内膜上皮细胞，直到第5代持续81天。此外，在冻融过程和顺序培养后，子宫内膜上皮和间质能够建立子宫内膜三维模型。我们在这里建立了体外培养的子宫内膜模型，作为难治性薄内膜的潜在治疗选择。通过细胞因子，膜结合分子，细胞外基质和间隙连接对子宫内膜上皮和基质细胞进行编排的三维培养模型将为探索植入现象的潜在机制提供新的框架。此外，改良的胚胎培养物，即所谓的“体外植入”，可能是生育治疗中的治疗方法。间隙连接将为探索植入现象的潜在机制提供一个新的框架。此外，改良的胚胎培养物，即所谓的“体外植入”，可能是生育治疗中的治疗方法。间隙连接将为探索植入现象的潜在机制提供一个新的框架。此外，改良的胚胎培养物，即所谓的“体外植入”，可能是生育治疗中的治疗方法。