Generation of inner ear cells from mouse iPS cells and its application to Age-related hearing loss

2016 Fiscal Year Final Research Report

• Project/Area Number: 15K20229
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Otorhinolaryngology
• Research Institution: Juntendo University
• Principal Investigator: FUKUNAGA Ichiro 順天堂大学, 医学(系)研究科(研究院), 博士研究員 (20746581)
• Research Collaborator: KAMIYA Kazusaku 順天堂大学, 医学部, 准教授 (10374159)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Keywords: iPS細胞 / 蝸牛支持細胞 / Connexin26 / ギャップ結合プラーク / 加齢性難聴 / 遺伝性難聴

Outline of Final Research Achievements

In this study, we generated Cx26 expressing cells with GJP formation from mouse iPS cells (iCx26GJC). To investigate whether the iCx26GJCs were functional, we performed scrape-loading assay with Lucifer yellow (LY) and Ca2+imaging.　In the iCx26GJC culture, we observed that LY diffused beyond the wounded parental cells. In contrast, such extent of dye transfer was not observed in undifferentiated iPSCs. Furthermore, iCx26GJC exhibited spontaneous Ca2+ transients and their propagation. The spontaneous Ca2+ signaling activity was clearly inhibited by the p2x receptor antagonist (PPADS) and connexin hemichannel blocker (FFA), and the activity was restored after removal of the inhibitors.
on the other hand, to enhance MSC invasion to cochlea tissue, we developed a novel transplant strategy by induction of SDF-1 /MCP-1 expression in host cochlear tissue and enhanced expression of their receptors, chemokine (C-C motif) receptor 2 (CCR2) and C-X-C chemokine receptor type 4 (CXCR4) in MSC.

Free Research Field

発生生物学