Graphene Oxide Hierarchical Patterns for the Derivation of Electrophysiologically Functional Neuron-like Cells from Human Neural Stem Cells.
Graphene has shown great potential for biomedical engineering applications due to its electrical conductivity, mechanical strength, flexibility, and biocompatibility. Topographical cues of culture substrates or tissue-engineering scaffolds regulate the behaviors and fate of stem cells. In this study, we developed a graphene oxide (GO)-based patterned substrate (GPS) with hierarchical structures capable of generating synergistic topographical stimulation to enhance integrin clustering, focal adhesion, and neuronal differentiation in human neural stem cells (hNSCs). The hierarchical structures of the GPS were composed of microgrooves (groove size: 5, 10, and 20 μm), ridges (height: 100-200 nm), and nanoroughness surfaces (height: ∼10 nm). hNSCs grown on the GPS exhibited highly elongated, aligned neurite extension along the ridge of the GPS and focal adhesion development that was enhanced compared to that of cells grown on GO-free flat substrates and GO substrates without the hierarchical structures. In particular, GPS with a groove width of 5 μm was found to be the most effective in activating focal adhesion signaling, such as the phosphorylation of focal adhesion kinase and paxillin, thereby improving neuronal lineage commitment. More importantly, electrophysiologically functional neuron-like cells exhibiting sodium channel currents and action potentials could be derived from hNSCs differentiated on the GPS even in the absence of any of the chemical agents typically required for neurogenesis. Our study demonstrates that GPS could be an effective culture platform for the generation of functional neuron-like cells from hNSCs, providing potent therapeutics for treating neurodegenerative diseases and neuronal disorders.
Department of Biotechnology, Yonsei University , Seoul 120-749, Republic of Korea.|Department of Biomaterials Science and Engineering, Yonsei University , Seoul 120-749, Republic of Korea.
Kisuk
K
Yang
School of Electrical and Electronic Engineering, Yonsei University , Seoul 120-749, Republic of Korea.
Jaehong
J
Lee
Department of Biotechnology, Yonsei University , Seoul 120-749, Republic of Korea.
Jong Seung
JS
Lee
School of Electrical and Electronic Engineering, Yonsei University , Seoul 120-749, Republic of Korea.
Dayeong
D
Kim
Department of Biotechnology, Yonsei University , Seoul 120-749, Republic of Korea.
Gyeong-Eon
GE
Chang
School of Electrical and Electronic Engineering, Yonsei University , Seoul 120-749, Republic of Korea.
Jungmok
J
Seo
Department of Biotechnology, Yonsei University , Seoul 120-749, Republic of Korea.
Eunji
E
Cheong
School of Electrical and Electronic Engineering, Yonsei University , Seoul 120-749, Republic of Korea.
Taeyoon
T
Lee
Department of Biotechnology, Yonsei University , Seoul 120-749, Republic of Korea.
Seung-Woo
SW
Cho
D002454:Cell Differentiation / Q000502:physiology; D022001:Focal Adhesions / Q000502:physiology; D006108:Graphite / Q000737:chemistry*; D006801:Humans; D058953:Neural Stem Cells / Q000166:cytology / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D010087:Oxides / Q000737:chemistry*; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods; D054457:Tissue Scaffolds
D010087:Oxides; D006108:Graphite
electrophysiology; focal adhesion; graphene oxide pattern; hierarchical topography; human neural stem cell; neuronal differentiation