Effective induction of inner ear hair cells from mouse embryonic stem cells

小鼠胚胎干细胞有效诱导内耳毛细胞

• 批准号: 22390318
• 负责人: YOSHIKAWA Masahide
• 金额: $ 11.98万
• 依托单位: Nara Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22390318/
• 关键词: 内耳; 再生医学; 有毛細胞; 幹細胞; 耳鼻科; 分化誘導; 聴覚再生; 細胞移植

Hearing loss is mainly caused by loss of sensory hair cells (HCs) in the organ of Corti or cochlea. Although embryonic stem (ES) cells are a promising source for cell therapy, little is known about the efficient generation of HC-like cells from ES cells. In the present study, we developed a single-medium culture method for growing embryoid bodies (EBs), in which conditioned medium (CM) from cultures of ST2 stromal cells (ST2-CM) was used for 14-day cultures of 4-day EBs. At the end of the 14-day cultures, up to 20% of the cells in EB outgrowths expressed HC-related markers, including Math1 (also known as Atoh1), myosin6, myosin7a, calretinin, α9AchR and Brn3c (also known as Pou4f3), and also showed formation of stereocilia-like structures. Further, we found that these cells were incorporated into the developing inner ear after transplantation into chick embryos. The present inner ear HC induction method using ST2-CM (HIST2 method) is quite simple and highly efficient to obtain ES-derived HC-like cells with a relatively short cultivation time.

听力损失主要是由 Corti 或耳蜗中的感觉毛细胞 (HC) 损失引起的。尽管胚胎干 (ES) 细胞是细胞治疗的一个有前景的来源，但人们对如何有效生成 HC 样细胞知之甚少。在本研究中，我们开发了一种用于生长胚状体 (EB) 的单一培养基培养方法，其中使用来自 ST2 基质细胞 (ST2-CM) 培养物的条件培养基 (CM) 培养 14 天。 4 天的 EB 培养结束时，EB 生长物中高达 20% 的细胞表达 HC 相关标记物，包括 Math1（也称为 Atoh1）、肌球蛋白 6、肌球蛋白 7a、钙视网膜蛋白、α9AchR 和Brn3c（也称为 Pou4f3），也显示出静纤毛样结构的形成，此外，我们发现这些细胞被纳入发育中的内部。目前使用 ST2-CM（HIST2 方法）的内耳 HC 诱导方法非常简单且高效，可以在相对较短的培养时间内获得 ES 衍生的 HC 样细胞。

项目成果

Efficient induction of inner ear hair cell-like cells from Math1-transfected mouse ES cells

从 Math1 转染的小鼠 ES 细胞中有效诱导内耳毛细胞样细胞

• 发表时间: 2013
• 作者: Yukiteru Ouji;Shigeaki Ishizaka;Fukumi Nakamura-Uchiyama;Akio Wanaka;Masahide Yoshikawa
• 通讯作者: Masahide Yoshikawa

In vitro differentiation of mouse embryonic stem cells into inner ear hair cell-like cells using stromal cells

使用基质细胞将小鼠胚胎干细胞体外分化为内耳毛细胞样细胞

• 发表时间: 2012
• 作者: Yukiteru Ouji;Shigeaki Ishizaka;Fukumi Nakamura-Uchiyama;Akio Wanaka;Masahide Yoshikawa
• 通讯作者: Masahide Yoshikawa

ES細胞から内耳有毛細胞への効率的分化誘導法の開発

开发ES细胞向内耳毛细胞高效分化诱导方法

• 发表时间: 2012
• 作者: 王寺幸輝;吉川正英
• 通讯作者: 吉川正英

Dermal Papilla Cells Serially Cultured With Wnt-10b Sustain Their Hair Follicle Induction Activity After Transplantation Into Nude Mice

• DOI: 10.3727/096368912x636867
• 发表时间: 2012-01-01
• 期刊: CELL TRANSPLANTATION
• 影响因子: 3.3
• 作者: Ouji, Yukiteru;Ishizaka, Shigeaki;Yoshikawa, Masahide
• 通讯作者: Yoshikawa, Masahide

In vitro differentiation of mouse embryonic stem cells into inner ear hair cell by stromal cells

小鼠胚胎干细胞通过基质细胞体外分化为内耳毛细胞

• 发表时间: 2010
• 作者: Ouji Y;Yoshikawa M;Ishizaka S.
• 通讯作者: Ishizaka S.