title: An Engineered N-Cadherin Substrate for Differentiation, Survival, and Selection of Pluripotent Stem Cell-Derived Neural Progenitors.
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.
authors.affiliation: Department of Biomolecular Engineering, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Amranul
authors.initials: A
authors.lastname: Haque
authors.affiliation: Department of Biomolecular Engineering, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Nihad
authors.initials: N
authors.lastname: Adnan
authors.affiliation: Department of Biomolecular Engineering, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Ali
authors.initials: A
authors.lastname: Motazedian
authors.affiliation: Department of Biological Information, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Farhima
authors.initials: F
authors.lastname: Akter
authors.affiliation: Department of Biomolecular Engineering, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Sharif
authors.initials: S
authors.lastname: Hossain
authors.affiliation: Department of Biomolecular Engineering, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Kutsuzawa
authors.affiliation: Department of Biomolecular Engineering, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Kakon
authors.initials: K
authors.lastname: Nag
authors.affiliation: Department of Biological Information, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Eiry
authors.initials: E
authors.lastname: Kobatake
authors.affiliation: Department of Biomolecular Engineering, Tokyo Institute of Technology, Yokohama, Japan.
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Akaike
mesh_terms: D000818:Animals; D015153:Blotting, Western; D015820:Cadherins / Q000235:genetics* / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics; D053595:Embryonic Stem Cells / Q000378:metabolism; D015870:Gene Expression; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D058953:Neural Stem Cells / Q000378:metabolism*; D016501:Neurites / Q000502:physiology; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059008:Time-Lapse Imaging; D051176:beta Catenin / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000378:metabolism; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: D015820:Cadherins; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins; D051176:beta Catenin; D054460:rho-Associated Kinases; D020741:rho GTP-Binding Proteins