Induction of differentiation of human embryonic stem cells into functional hair-cell-like cells in the absence of stromal cells

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• 作者：Jie Ding^a ; ^b ; Zihua Tang^a ; Jiarong Chen^a ; Haosong Shi^c ; Jianling Chen^a ; Cuicui Wang^a ; Cui Zhang^a ; Liang Li^a ; Ping Chen^a ; ^d ; Jinfu Wang^a ; ^{wjfu@zju.edu.cn}
• 关键词：hESCs ; human embryonic stem cells ; OEPs ; otic epithelial progenitors ; ONPs ; otic neural progenitors ; FGF3 ; fibroblast growth factor 3 ; FGF10 ; fibroblast growth factor 10 ; EGF ; epidermal growth factor ; bFGF ; basic fibroblast growth factor ; RA ; all-trans retinoic acid ; MEF ; mouse embryonic fibroblast ; KOSR ; knockout serum replacement ; RT-PCR ; reverse transcription-polymerase chain reaction ; DAPI ; 4&prime ; 6-diamidino-2-phenylindole ; SEM ; scanning electron microscopy
• 刊名：The International Journal of Biochemistry & Cell Biology
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：81
• 期：part_PA
• 页码：208-222
• 全文大小：5534 K
• 卷排序：81

文摘

Sensorineural hearing loss and vestibular dysfunction have become the most common forms of sensory defects. Stem cell-based therapeutic strategies for curing hearing loss are being developed. Several attempts to develop hair cells by using chicken utricle stromal cells as feeder cells have resulted in phenotypic conversion of stem cells into inner ear hair-cell-like cells. Here, we induced the differentiation of human embryonic stem cells (hESCs) into otic epithelial progenitors (OEPs), and further induced the differentiation of OEPs into hair-cell-like cells using different substrates. Our results showed that OEPs cultured on the chicken utricle stromal cells with the induction medium could differentiate into hair-cell-like cells with stereociliary bundles. Co-culture with stromal cells, however, may be problematic for subsequent examination of the induced hair-cell-like cells. In order to avoid the interference from stromal cells, we cultured OEPs on laminin with different induction media and examined the effects of the induction medium on the differentiation potentials of OEPs into hair-cell-like cells. The results revealed that the culture of OEPs on laminin with the conditioned medium from chicken utricle stromal cells supplemented with EGF and all-trans retinoic acid (RA) could promote the organization of cells into epithelial clusters displaying hair-cell-like cells with stereociliary bundles. These cells also displayed the expected electrophysiological properties.