| RRC ID |
52289
|
| 著者 |
Ouji Y, Sakagami M, Omori H, Higashiyama S, Kawai N, Kitahara T, Wanaka A, Yoshikawa M.
|
| タイトル |
Efficient induction of inner ear hair cell-like cells from mouse ES cells using combination of Math1 transfection and conditioned medium from ST2 stromal cells.
|
| ジャーナル |
Stem Cell Res
|
| Abstract |
We sought to establish a more efficient technique for induction of inner ear hair cell-like cells (HC-like cells) from embryonic stem cells (ES cells) by using a combination of two previously reported methods; ST2 stromal cell-conditioned medium, known to be favorable for HC-like cell induction (HIST2 method), and ES cells with transfer of the Math1 gene (Math1-ES cells). Math1-ES cells carrying Tet-inducible Math1 were cultured for 14days with doxycycline in conditioned medium from cultures of ST2 stromal cells following formation of 4-day embryoid bodies (EBs). Although each of the previously introduced methods have been reported to induce approximately 20% HC-like cells and 10% HC-like cells in their respective populations in EB outgrowths at the end of the culture periods, the present combined method was able to generate approximately 30% HC-like cells expressing HC-related markers (myosin6, myosin7a, calretinin, α9AchR, Brn3c), which showed remarkable formation of stereocilia-like structures. Analysis of expressions of marker genes specific for cochlear (Lmod3, Emcn) and vestibular (Dnah5, Ptgds) cells indicated that our HIST2 method may lead to induction of cochlear- and vestibular-type cells. In addition, continuous Math1 induction by doxycycline without use of the HIST2 method preferentially induced cochlear markers with negligible effects on vestibular marker induction.
|
| 巻・号 |
23
|
| ページ |
50-56
|
| 公開日 |
2017-8-1
|
| DOI |
10.1016/j.scr.2017.06.013
|
| PII |
S1873-5061(17)30121-6
|
| PMID |
28689068
|
| MeSH |
Animals
Basic Helix-Loop-Helix Transcription Factors / genetics*
Basic Helix-Loop-Helix Transcription Factors / metabolism
Biomarkers / metabolism
Cell Line
Cells, Cultured
Cochlea / cytology
Culture Media, Conditioned / pharmacology*
Embryoid Bodies / cytology
Embryoid Bodies / drug effects
Gene Expression Regulation / drug effects
Hair Cells, Auditory, Inner / cytology*
Mechanotransduction, Cellular
Mice
Mouse Embryonic Stem Cells / cytology*
Myosins / metabolism
Stereocilia / metabolism
Stromal Cells / metabolism
Transfection*
Vestibule, Labyrinth / cytology
|
| IF |
4.495
|
| 引用数 |
5
|
| リソース情報 |
| ヒト・動物細胞 |
ST2(RCB0224) |
