In Vitro Epithelial Differentiation of Human Induced Pluripotent Stem Cells for Vocal Fold Tissue Engineering

被引:9
|
作者
Imaizumi, Mitsuyoshi [1 ,3 ]
Sato, Yuka [1 ]
Yang, David T. [2 ]
Thibeault, Susan L. [1 ]
机构
[1] Univ Wisconsin, Div Otolaryngol Head & Neck Surg, Madison, WI 53705 USA
[2] Univ Wisconsin, Dept Pathol & Lab Med, Madison, WI 53705 USA
[3] Fukushima Med Univ, Dept Otolaryngol, Fukushima, Japan
来源
ANNALS OF OTOLOGY RHINOLOGY AND LARYNGOLOGY | 2013年 / 122卷 / 12期
关键词
hiPS; hyaluronic acid hydrogel; induced pluripotent stem cell; tissue engineering; vocal fold; SYNTHETIC EXTRACELLULAR-MATRIX; EPIDERMAL-GROWTH-FACTOR; REGENERATION; PROLIFERATION; HYALURONAN; VISCOELASTICITY; FIBROBLASTS; INJECTION;
D O I
10.1177/000348941312201203
中图分类号
R76 [耳鼻咽喉科学];
学科分类号
100213 ;
摘要
Objectives: We determined the feasibility and optimization of differentiating human induced pluripotent stem cells (hiPS) into nonkeratinized stratified squamous epithelial cells for vocal fold engineering. Methods: hiPS were cultured and assessed for differentiation in 3 conditions: a 3-dimensional (3D) hyaluronic acid (HA) hydrogel scaffold, a 3D HA hydrogel scaffold with epidermal growth factor (EGF), and a 3D HA hydrogel scaffold co-cultured with human vocal fold fibroblasts (hVFF). After 1, 2, and 4 weeks of cultivation, hiPS were selected for histology, immunohistochemistry, and/or transcript expression analysis. Results: At 4 weeks, hiPS cultivated with hVFF or with EGF had significantly decreased levels of Oct 3/4, indicating loss of pluripotency. Immunofluorescence revealed the presence of pancytokeratin and of cytokeratin (CK) 13 and 14 epithelial-associated proteins at 4 weeks after cultivation in hiPS EGF and hiPS hVFF cultures. The transcript expression level of CK14 was significantly increased for hiPS hVFF cultures only and was measured concomitantly with cell morphology that was clearly cohesive and displayed a degree of nuclear polarity suggestive of epithelial differentiation. Conclusions: We found that hiPS cultivated in 3D HA hydrogel with hVFF demonstrated the most robust conversion evidence to date of epithelial differentiation. Further work is necessary to focus on amplification of these progenitors for application in vocal fold regenerative biology.
引用
收藏
页码:737 / 747
页数:11
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