A recombinant extracellular matrix of N-cadherin enhances generation and selection of pluripotent stem cell-derived neurons via suppression of Rho kinase.
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1
Tokyo Institute of Technology, Biomolecular Engineering, Japan
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2
University of California, Biomedical Engineering, United States
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3
Foundation for Advancement of International Science, Biomaterials Center for Regenerative Medical Engineering, Japan
The formation of appropriate neural circuits depends on coordinated interactions between extrinsic guiding cues and intracellular signaling, which are often mediated by cell adhesion molecules such as N-cadherin. In this study, we demonstrated that embryonic stem cell (ESC) and induced pluripotent stem cell (iPSC)-derived neural progenitor cells (NPCs) extend longer neurites in response to matrix-anchored N-cadherin, when compared to controls. Although exogenous growth factors such as CKI-7 and SB43152 n-hydrate induced more robust NPC marker expression in ESCs and iPSCs on day 5, further maturation towards functional neural cells was similar in ESC and iPSCs progeny cultured on N-cadherin substrate, irrespective of the addition of extrinsic neuroinductive factors. The effects of N-cadherin on neurite outgrowth was significantly reduced in the presence of RhoA activator or N-cadherin neutralizing antibody without any noticeable changes in the presence of soluble cadherin homophilic ligands or inhibitor to fibroblast growth factor receptor (PD173074). In addition, the expression of β-catenin was reduced in NPCs cultured on N-cadherin surface, with lowest expression was observed in dissociated single-cells.To generate homogeneous cell populations, we described a robust protocol to purge undifferentiated ESCs and iPSCs from heterogeneous NPCs by N-cadherin-based homophilic adhesion. Using this label-free cell selection approach we enriched viable differentiated neurons plated as single-cells 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.
This work was funded by Japan Society for the Promotion of Science (S) No.23220014.
References:
[1] Haque A, Yue XS, Motazedian A, Tagawa Y, Akaike T. Characterization and neural differentiation of mouse embryonic and induced pluripotent stem cells on cadherin-based substrata. Biomaterials. 2012. 33: p. 5094-5106.
[2] Haque A, Adnan N, Motazedian A, Akter F, Hossain S, Kutsuzawa K, et al. An engineered N-cadherin substrate for differentiation, survival, and selection of pluripotent stem cell-derived neural progenitors. PLoS One. 2015. 10: e0135170.
Keywords:
Cell Differentiation,
Extracellular Matrix,
Regenerative Medicine,
stem cell
Conference:
10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.
Presentation Type:
Poster
Topic:
Synthetic scaffolds as extracellular matrices
Citation:
Haque
A,
Motazedian
A,
Adnan
N and
Akaike
T
(2016). A recombinant extracellular matrix of N-cadherin enhances generation and selection of pluripotent stem cell-derived neurons via suppression of Rho kinase..
Front. Bioeng. Biotechnol.
Conference Abstract:
10th World Biomaterials Congress.
doi: 10.3389/conf.FBIOE.2016.01.00117
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Received:
27 Mar 2016;
Published Online:
30 Mar 2016.