title: Combinatorial protein display for the cell-based screening of biomaterials that direct neural stem cell differentiation.
abstract: Neural stem cell (NSC) has emerged as a potential source for cell replacement therapy following traumatic injuries and degenerative diseases of the central nervous system. However, clinical applications of NSC further require technological advances especially for controlling differentiation of NSC. This study aimed at developing biomaterials that serve to expand undifferentiated NSC or to induce cells with specific phenotypes. Our approach is to construct composite biomaterials that consist of extracellular matrix components and growth factors. In order to optimize matrix-growth factor combinations, we conducted the parallel and rapid screening of composite biomaterials through assays using cell-based arrays. The photo-assisted patterning of an alkanethiol self-assembled monolayer was employed to achieve site-addressable combinatorial immobilization of natural and synthetic matrices incorporated with growth factors including epidermal growth factor (EGF), ciliary neurotrophic factor (CNTF), nerve growth factor (NGF), and neurotrophin-3 (NT-3). NSC obtained from the rat embryonic striatum was cultured directly on the array to screen for cell adhesion, proliferation, and promotion of neuronal and glial specification. The results showed that the significant number of cells adhered to laminin-1, fibronectin, ProNectin, and poly(ethyleneimine). It was found that cells proliferated most extensively on a spot with immobilized EGF among the spots with different matrix-growth factor combinations. The results also showed that neuronal differentiation was promoted on the spots with immobilized NGF or NT-3, and astroglial differentiation with CNTF. Importantly, observed effects of growth factors were frequently altered depending on the type of co-immobilized matrices, suggesting synergic effects of adhesion and growth factor signals.
authors.affiliation: Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Nakajima
authors.forename: Toshinari
authors.initials: T
authors.lastname: Ishimuro
authors.forename: Koichi
authors.initials: K
authors.lastname: Kato
authors.forename: In-Kap
authors.initials: IK
authors.lastname: Ko
authors.forename: Isao
authors.initials: I
authors.lastname: Hirata
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Arima
authors.forename: Hiroo
authors.initials: H
authors.lastname: Iwata
mesh_terms: D000327:Adsorption; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D020650:Combinatorial Chemistry Techniques; D008422:Materials Testing / Q000379:methods*; D009414:Nerve Growth Factors / Q000008:administration & dosage* / Q000737:chemistry; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D009474:Neurons / Q000166:cytology* / Q000187:drug effects; D051381:Rats; D011916:Rats, Inbred F344; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D001672:Biocompatible Materials; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins