1 Department of Medical Oncology and Niigata University Graduate School of Medical and Dental Sciences , Niigata, Japan .
2 Department of Thoracic Surgery, Niigata University Graduate School of Medical and Dental Sciences , Niigata, Japan .
Tissue Eng Part C Methods. 2018 Sep;24(9):524-533. doi: 10.1089/ten.TEC.2018.0115.
The outcomes of tracheal transplantation for the treatment of airway stenosis are unsatisfactory. We investigated the feasibility of regeneration of the trachea using a rat decellularized tracheal scaffold and mouse-induced pluripotent stem (iPS) cells for in vivo transplantation. The rat trachea was first decellularized using a detergent/enzymatic treatment method. We successfully established a centrifugation method that can transplant cells onto the luminal surface of the decellularized rat tracheal scaffold circumferentially. Two types of mouse iPS cells were differentiated into definitive endoderm cells and transplanted onto the luminal surface of the decellularized tracheal matrix scaffold using this centrifugation method. For in vivo study, normal rat tracheas, no-cell rat tracheal scaffolds, or rat tracheal scaffolds recellularized with rat tracheal epithelial cells (EGV-4T) were orthotopically transplanted on F344 rats, and rat tracheal scaffolds recellularized with mouse iPS cells were transplanted on F344/NJc1-rnu/rnu rats. Rats transplanted with no-cell scaffolds or scaffolds recellularized with EGV-4T survived for 1 month, although airway stenosis was observed. One of the F344/NJc1-rnu/rnu rats transplanted with rat trachea regenerated using mouse iPS cells survived over 5 weeks. Histological analysis indicated the cause of death was airway stenosis due to colonic cellular proliferation of undifferentiated iPS cells. Re-epithelialization with numerous ciliated epithelial cells was observed in one of the rats transplanted with trachea bioengineered using iPS cells. In this study, we present a simple and efficient tracheal tissue engineering model using a centrifugation method in a small-animal model. Tissue-engineered trachea using decellularized tracheal scaffolds and iPS cells is potentially applicable for tracheal transplantation.
气管移植治疗气道狭窄的效果并不理想。我们研究了使用大鼠脱细胞气管支架和小鼠诱导多能干细胞（iPS 细胞）进行体内移植来实现气管再生的可行性。首先，我们使用去污剂/酶处理方法对大鼠气管进行脱细胞处理。我们成功建立了一种离心方法，可以将细胞移植到脱细胞大鼠气管支架的腔面表面上。我们使用这种离心方法将两种类型的小鼠 iPS 细胞分化为确定的内胚层细胞，并将其移植到脱细胞气管基质支架的腔面表面上。在体内研究中，我们将正常大鼠气管、无细胞大鼠气管支架或用大鼠气管上皮细胞（EGV-4T）再细胞化的大鼠气管支架原位移植到 F344 大鼠体内，并用小鼠 iPS 细胞再细胞化的大鼠气管支架移植到 F344/NJc1-rnu/rnu 大鼠体内。移植无细胞支架或用 EGV-4T 再细胞化的支架的大鼠存活了 1 个月，但观察到气道狭窄。用小鼠 iPS 细胞再生的大鼠气管移植的 1 只 F344/NJc1-rnu/rnu 大鼠存活了 5 周以上。组织学分析表明，死亡的原因是由于未分化的 iPS 细胞的结直肠细胞增殖导致气道狭窄。用 iPS 细胞构建的气管之一移植的大鼠观察到有大量纤毛上皮细胞再上皮化。在本研究中，我们在小动物模型中使用离心方法建立了一种简单有效的气管组织工程模型。使用脱细胞气管支架和 iPS 细胞构建的组织工程气管有望应用于气管移植。
