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Development of A 3D Stem Cell-derived Neuron Model for Neuronal Drug Testing
Liu Qingxi1, Wu Yueting1,2, Sun He1, Cui Zhanfeng3, Li Zhaohui3*
1Tianjin Weikai Bioeng Ltd., Tianjin 300457, China; 2Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; 3Institute of Biomedical Engineering, Oxford University, Oxford OX13PJ, UK
Abstract: The neural stem cells were cultured in hyaluronic acid-based three-dimensional (3D) scaffold and induced differentiation for 1， 7 and 14 days， besides the conventional two-dimensional (2D) culture was set as a control. The expressions of stemness marker (nestin)， neuronal marker (tubulin) and glial cell marker (glial fibrillary acidic protein， GFAP) were detected using immunocytochemistry staining and Real-time PCR; The cell survival and proliferation ability during differentiation culture and drug testing were assayed using the CCK-8 Kit and Live-Dead Cell Staining Kit. During culture period， tubulin expression was significantly increased and glial cell marker GFAP expression reduced， whereas the changes observed in 3D model were more obvious. Cells cultured in the 3D model were shown a significant protective effect on drug-induced nerve damage post differentiation. Neural stem cells cultured in 3D hydrogel retained a high differentiation potential to form neuron cells compared to conventional 2D culture. The 3D culture model could provide an additional protection on drug-induced nerve damage post differentiation， also the developed 3D stem cell-derived neuronal model could be an advanced in-vitro model for drug toxicity testing and for efficacy and safety assessments in stem cell therapy to treat neurodegenerative diseases.
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