Journal
BIOMEDICINES
Volume 10, Issue 7, Pages -Publisher
MDPI
DOI: 10.3390/biomedicines10071534
Keywords
optogenetics; channelrhodopsin-2 (ChR2); neuronal differentiation; SH-SY5Y cells; RGD-alginate; hydrogels; 3D culture; neurodegenerative disease
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Funding
- Ministry of Higher Education, Malaysia
- Biotechnology and Biological Sciences Research Council (BBSRC), United Kingdom [BB/H008527/1]
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This study proposed a 3D platform based on RGD-alginate for encapsulating optogenetically modified human neuronal cells, demonstrating their survival and ability to express functional neurons.
The cases of brain degenerative disease will rise as the human population ages. Current treatments have a transient effect and lack an investigative system that is physiologically relevant for testing. There is evidence suggesting optogenetic stimulation is a potential strategy; however, an in vitro disease and optogenetic model requires a three-dimensional microenvironment. Alginate is a promising material for tissue and optogenetic engineering. Although it is bioinert, alginate hydrogel is transparent and therefore allows optical penetration for stimulation. In this study, alginate was functionalized with arginine-glycine-aspartate acid (RGD) to serve as a 3D platform for encapsulation of human SH-SY5Y cells, which were optogenetically modified and characterized. The RGD-alginate hydrogels were tested for swelling and degradation. Prior to encapsulation, the cells were assessed for neuronal expression and optical-stimulation response. The results showed that RGD-alginate possessed a consistent swelling ratio of 18% on day 7, and degradation remained between 3.7-5% throughout 14 days. Optogenetically modified SH-SY5Y cells were highly viable (>85%) after lentiviral transduction and neuronal differentiation. The cells demonstrated properties of functional neurons, developing beta III tubulin (TuJ1)-positive long neurites, forming neural networks, and expressing vGlut2. Action potentials were produced upon optical stimulation. The neurons derived from human SH-SY5Y cells were successfully genetically modified and encapsulated; they survived and expressed ChR2 in an RGD-alginate hydrogel system.
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