Uterus tissue engineering may dismantle limitations in current uterus transplantation protocols. A uterine biomaterial populated with patient-derived cells could potentially serve as a graft to circumvent complicated surgery of live donors, immunosuppressive medication and rejection episodes. Repeated uterine bioengineering studies on rodents have shown promising results using decellularised scaffolds to restore fertility in a partially impaired uterus and now mandate experiments on larger and more human-like animal models. The aim of the presented studies was therefore to establish adequate protocols for scaffold generation and prepare for future in vivo sheep uterus bioengineering experiments. Three decellularisation protocols were developed using vascular perfusion through the uterine artery of whole sheep uteri obtained from slaughterhouse material. Decellularisation solutions used were based on 0.5% sodium dodecyl sulphate (Protocol 1) or 2% sodium deoxycholate (Protocol 2) or with a sequential perfusion of 2% sodium deoxycholate and 1% Triton X-100 (Protocol 3). The scaffolds were examined by histology, extracellular matrix quantification, evaluation of mechanical properties and the ability to support foetal sheep stem cells after recellularisation. We showed that a sheep uterus can successfully be decellularised while maintaining a high integrity of the extracellular components. Uteri perfused with sodium deoxycholate (Protocol 2) were the most favourable treatment in our study based on quantifications. However, all scaffolds supported stem cells for 2 weeks in vitro and showed no cytotoxicity signs. Cells continued to express markers for proliferation and maintained their undifferentiated phenotype. Hence, this study reports three valuable decellularisation protocols for future in vivo sheep uterus bioengineering experiments.

中文翻译：

---

走向子宫组织工程：绵羊子宫脱细胞的比较研究。

子宫组织工程可以消除当前子宫移植方案中的局限性。一种由患者来源的细胞组成的子宫生物材料有可能充当移植物，以规避活体供体的复杂手术，免疫抑制药物治疗和排斥反应。对啮齿类动物的重复子宫生物工程研究显示，使用脱细胞支架在部分受损的子宫中恢复生育力的结果令人鼓舞，现在需要在更大，更像人类的动物模型上进行实验。因此，本研究的目的是建立足够的支架生成方案，并为将来的体内绵羊子宫生物工程实验做准备。利用从屠宰场获得的全羊子宫的子宫动脉血管灌注，开发了三种脱细胞方案。所用的脱细胞溶液是基于0.5％的十二烷基硫酸钠（协议1）或2％的脱氧胆酸钠（协议2），或依次灌注2％的脱氧胆酸钠和1％的Triton X-100（协议3）。通过组织学，细胞外基质定量，机械性能评估和重新细胞化后支持胎羊干细胞的能力来检查支架。我们表明绵羊子宫可以成功地脱细胞，同时保持细胞外成分的高度完整性。根据定量研究，灌注脱氧胆酸钠（方案2）的子宫是我们研究中最有利的治疗方法。然而，所有支架在体外均支持干细胞2周，并且未显示任何细胞毒性迹象。细胞继续表达增殖标记并保持其未分化表型。因此，本研究报告了三种有价值的脱细胞方案，可用于未来的体内绵羊子宫生物工程实验。