Ikeda Masakazu, Imaizumi Mitsuyoshi, Yoshie Susumu, Nakamura Ryosuke, Otsuki Koshi, Murono Shigeyuki, Omori Koichi
1 Department of Otolaryngology, Fukushima Medical University, Fukushima, Japan.
Ann Otol Rhinol Laryngol. 2017 Jul;126(7):517-524. doi: 10.1177/0003489417713504.
Compared with using autologous tissue, the use of artificial materials in the regeneration of tracheal defects is minimally invasive. However, this technique requires early epithelialization on the inner side of the artificial trachea. After differentiation from induced pluripotent stem cells (iPSCs), tracheal epithelial tissues may be used to produce artificial tracheas. Herein, we aimed to demonstrate that after differentiation from fluorescent protein-labeled iPSCs, tracheal epithelial tissues survived in nude rats with tracheal defects.
Red fluorescent tdTomato protein was electroporated into mouse iPSCs to produce tdTomato-labeled iPSCs. Embryoid bodies derived from these iPSCs were then cultured in differentiation medium supplemented with growth factors, followed by culture on air-liquid interfaces for further differentiation into tracheal epithelium. The cells were implanted with artificial tracheas into nude rats with tracheal defects on day 26 of cultivation. On day 7 after implantation, the tracheas were exposed and examined histologically.
Tracheal epithelial tissue derived from tdTomato-labeled iPSCs survived in the tracheal defects. Moreover, immunochemical analyses showed that differentiated tissues had epithelial structures similar to those of proximal tracheal tissues.
After differentiation from iPSCs, tracheal epithelial tissues survived in rat bodies, warranting the use of iPSCs for epithelial regeneration in tracheal defects.
与使用自体组织相比，在气管缺损再生中使用人工材料的侵入性最小。然而，该技术要求人工气管内侧早期上皮化。从诱导多能干细胞（iPSC）分化后，气管上皮组织可用于制造人工气管。在此，我们旨在证明荧光蛋白标记的iPSC分化后，气管上皮组织在有气管缺损的裸鼠体内存活。
将红色荧光tdTomato蛋白电穿孔导入小鼠iPSC以产生tdTomato标记的iPSC。然后将这些iPSC来源的胚状体在补充有生长因子的分化培养基中培养，随后在气液界面上培养以进一步分化为气管上皮。在培养第26天，将细胞与人工气管一起植入有气管缺损的裸鼠体内。植入后第7天，暴露气管并进行组织学检查。
源自tdTomato标记的iPSC的气管上皮组织在气管缺损处存活。此外，免疫化学分析表明，分化组织具有与近端气管组织相似的上皮结构。
iPSC分化后，气管上皮组织在大鼠体内存活，这为在气管缺损中使用iPSC进行上皮再生提供了依据。
