Original articles
Cheng Chen, Xin Dong, Kai-Heng Fang, Fang Yuan, Yao Hu, Min Xu, Yu Huang, Xixiang Zhang, Danjun Fang, Yan Liu. Develop a 3D neurological disease model of human cortical glutamatergic neurons using micropillar-based scaffolds[J]. Acta Pharmaceutica Sinica B, 2019, 9(3): 557-564

Develop a 3D neurological disease model of human cortical glutamatergic neurons using micropillar-based scaffolds
Cheng Chena,b,c, Xin Donga,b, Kai-Heng Fanga,b, Fang Yuana,b, Yao Hua,b, Min Xua,b, Yu Huangc, Xixiang Zhangd, Danjun Fangb, Yan Liua,b,e
a Institute for Stem Cell and Neural Regeneration, School of Pharmacy, Nanjing Medical University, Nanjing 211166, China;
b Key Laboratory of Targeted Intervention of Cardiovascular Disease, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, School of Pharmacy, Nanjing Medical University, Nanjing 211166, China;
c Department of Biological Engineering, Utah State University, Logan, UT 84322, USA;
d Physical Sciences and Engineering Division(PSE), King Abdullah University of Science and Technology, Thuwal 23955-6900, Kingdom of Saudi Arabia;
e Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China
Establishing an effective three-dimensional (3D) in vitro culture system to better model human neurological diseases is desirable, since the human brain is a 3D structure. Here, we demonstrated the development of a polydimethylsiloxane (PDMS) pillar-based 3D scaffold that mimicked the 3D microenvironment of the brain. We utilized this scaffold for the growth of human cortical glutamatergic neurons that were differentiated from human pluripotent stem cells. In comparison with the 2D culture, we demonstrated that the developed 3D culture promoted the maturation of human cortical glutamatergic neurons by showing significantly more MAP2 and less Ki67 expression. Based on this 3D culture system, we further developed an in vitro disease-like model of traumatic brain injury (TBI), which showed a robust increase of glutamate-release from the neurons, in response to mechanical impacts, recapitulating the critical pathology of TBI. The increased glutamate-release from our 3D culture model was attenuated by the treatment of neural protective drugs, memantine or nimodipine. The established 3D in vitro human neural culture system and TBI-like model may be used to facilitate mechanistic studies and drug screening for neurotrauma or other neurological diseases.
Key words:    3D culture    Cortical glutamatergic neurons    Human pluripotent stem cells    Cell differentiation    Disease modeling    Traumatic brain injury    Neural protective drugs    Drug screening   
Received: 2018-12-24     Revised:
DOI: 10.1016/j.apsb.2019.03.004
Funds: This study was supported by the Strategic Priority Research Program of the Chinese Academy of Sciences (Grant No. XDA16010306), the National Natural Science Foundation of China Grants (91849117 and 81471301), Key Research and Development Program of China (2016YFC1306703), The National Jiangsu Outstanding Young Investigator Program (BK20160044, China), Jiangsu Province's Innovation Person (China), the Natural Science Foundation of the Jiangsu Higher Education Institutions of China Project (Grant No. 17KJB180010).
Corresponding author: Danjun Fang, Yan Liu     Email:djf@njmu.edu.cn;yanliu@njmu.edu.cn
Author description:
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Cheng Chen
Xin Dong
Kai-Heng Fang
Fang Yuan
Yao Hu
Min Xu
Yu Huang
Xixiang Zhang
Danjun Fang
Yan Liu

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