Evaluation of the Use of Induced Pluripotent Stem Cells (iPSCs) for the Regeneration of Tracheal Cartilage
Evaluation of the Use of Induced Pluripotent Stem Cells (iPSCs) for the Regeneration of Tracheal Cartilage
复制标题
DOI：
10.3727/096368912x653147
复制
发表时间：
2013-01-01
影响因子：
3.3
通讯作者：
Omori, Koichi
中科院分区：
文献类型：
--
作者：
Imaizumi, Mitsuyoshi;Nomoto, Yukio;Omori, Koichi
喉气管狭窄的治疗仍然是一个挑战，因为治疗往往需要多阶段的程序，成功的拔管有时失败后，一系列的操作。诱导多能干细胞（iPSC）于2006年产生。这些细胞能够无限对称的自我更新，从而为组织工程应用提供了无限的细胞来源。我们之前已经报道了使用含有iPSCs的三维（3D）支架进行气管壁再生。然而，分化成软骨的效率低。此外，无法证明软骨组织实际上来自植入的iPSC。本研究的目的是评估和改善iPSCs用于气管软骨再生的使用。在软骨细胞分化培养基中的3D支架中体外培养iPSC。培养后，检查向软骨细胞的分化。流式细胞仪分析未分化细胞比例。在24只裸大鼠的气管缺损处植入三维支架作为损伤部位。在体外分化成软骨细胞，证实组织学，表型和遗传。流式细胞仪分析表明，未分化细胞的数量减少。在植入诱导的iPSC的11只大鼠中的6只中，在再生的气管壁中观察到软骨组织，但在植入对照和未诱导的iPSC的13只大鼠中没有观察到软骨组织。软骨特异性蛋白质的表达也在含有iPSC的3D支架中体内证实。通过激光显微切割（LMD）和聚合酶链反应（PCR）技术的组合，在软骨组织样品中证实了源自iPSC的GFP基因的存在。我们的研究表明，iPSCs具有体外分化为软骨细胞的潜力。软骨组织在体内再生。我们的研究结果表明，诱导多能干细胞可能成为气管软骨再生的新细胞来源。
The treatment of laryngotracheal stenosis remains a challenge as treatment often requires multistaged procedures, and successful decannulation sometimes fails after a series of operations. Induced pluripotent stem cells (iPSCs) were generated in 2006. These cells are capable of unlimited symmetrical self-renewal, thus providing an unlimited cell source for tissue-engineering applications. We have previously reported tracheal wall regeneration using a three-dimensional (3D) scaffold containing iPSCs. However, the efficiency of differentiation into cartilage was low. In addition, it could not be proven that the cartilage tissues were in fact derived from the implanted iPSCs. The purpose of this study was to evaluate and improve the use of iPSCs for the regeneration of tracheal cartilage. iPSCs were cultured in vitro in a 3D scaffold in chondrocyte differentiation medium. After cultivation, differentiation into chondrocytes was examined. The ratio of undifferentiated cells was analyzed by flow cytometry. The 3D scaffolds were implanted into tracheal defects, as an injury site, in 24 nude rats. Differentiation into chondrocytes in vitro was confirmed histologically, phenotypically, and genetically. Flow cytometric analysis demonstrated that the population of undifferentiated cells was decreased. Cartilage tissue was observed in the regenerated tracheal wall in 6 of 11 rats implanted with induced iPSCs, but in none of 13 rats implanted with the control and noninduced iPSCs. The expression of cartilage-specific protein was also demonstrated in vivo in 3D scaffolds containing iPSCs. The presence of the GFP gene derived from iPSCs was confirmed in samples of cartilage tissue by the combination of laser microdissection (LMD) and polymerase chain reaction (PCR) techniques. Our study demonstrated that iPSCs have the potential to differentiate into chondrogenic cells in vitro. Cartilage tissue was regenerated in vivo. Our results suggest that iPSCs could be a new cell source for the regeneration of tracheal cartilage.