| RRC ID |
45477
|
| 著者 |
Haque A, Adnan N, Motazedian A, Akter F, Hossain S, Kutsuzawa K, Nag K, Kobatake E, Akaike T.
|
| タイトル |
An Engineered N-Cadherin Substrate for Differentiation, Survival, and Selection of Pluripotent Stem Cell-Derived Neural Progenitors.
|
| ジャーナル |
PLoS One
|
| Abstract |
For stem cell-based treatment of neurodegenerative diseases a better understanding of key developmental signaling pathways and robust techniques for producing neurons with highest homogeneity are required. In this study, we demonstrate a method using N-cadherin-based biomimetic substrate to promote the differentiation of mouse embryonic stem cell (ESC)- and induced pluripotent stem cell (iPSC)-derived neural progenitor cells (NPCs) without exogenous neuro-inductive signals. We showed that substrate-dependent activation of N-cadherin reduces Rho/ROCK activation and β-catenin expression, leading to the stimulation of neurite outgrowth and conversion into cells expressing neural/glial markers. Besides, plating dissociated cells on N-cadherin substrate can significantly increase the differentiation yield via suppression of dissociation-induced Rho/ROCK-mediated apoptosis. Because undifferentiated ESCs and iPSCs have low affinity to N-cadherin, plating dissociated cells on N-cadherin-coated substrate increase the homogeneity of differentiation by purging ESCs and iPSCs (~30%) from a mixture of undifferentiated cells with NPCs. Using this label-free cell selection approach we enriched differentiated NPCs plated as monolayer without ROCK inhibitor. Therefore, N-cadherin biomimetic substrate provide a powerful tool for basic study of cell-material interaction in a spatially defined and substrate-dependent manner. Collectively, our approach is efficient, robust and cost effective to produce large quantities of differentiated cells with highest homogeneity and applicable to use with other types of cells.
|
| 巻・号 |
10(8)
|
| ページ |
e0135170
|
| 公開日 |
2015-1-1
|
| DOI |
10.1371/journal.pone.0135170
|
| PII |
PONE-D-15-25411
|
| PMID |
26244942
|
| PMC |
PMC4526632
|
| MeSH |
Animals
Blotting, Western
Cadherins / genetics*
Cadherins / metabolism
Cell Culture Techniques / methods
Cell Differentiation / genetics*
Cell Line
Cell Survival / genetics
Embryonic Stem Cells / metabolism
Gene Expression
Immunoglobulin Fc Fragments / genetics
Immunoglobulin Fc Fragments / metabolism
Induced Pluripotent Stem Cells / metabolism
Mice
Microscopy, Confocal
Neural Stem Cells / metabolism*
Neurites / physiology
Pluripotent Stem Cells / metabolism*
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Time-Lapse Imaging
beta Catenin / metabolism
rho GTP-Binding Proteins / metabolism
rho-Associated Kinases / metabolism
|
| IF |
2.74
|
| 引用数 |
10
|
|
WOS 分野
|
CELL BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
iPS-MEF-Ng-20D-17(APS0001) |
