Construction of hiPSC-derived Excitatory Neural Network-like Tissue
10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.20230529.001
- VernacularTitle:兴奋性hiPSC源性类神经网络组织的构建
- Author:
Qi-song SU
1
;
Ge LI
2
;
Jin-hai XU
3
;
Bin JIANG
3
;
Qing-shuai WEI
2
;
Xiang ZENG
2
;
Yuan-shan ZENG
2
;
Ping ZHU
1
Author Information
1. School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China
2. Department of Histology and Embryology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China
3. Guangdong Province Key Laboratory of Brain Function and Disease, Guangzhou 510080, China
- Publication Type:Journal Article
- Keywords:
hiPSC-NPCs;
DON;
synapse;
excitatory neurons;
neural network-like tissue
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2023;44(4):625-633
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo construct a neural network-like tissue with the potential of synaptic formation in vitro by seeding human induced pluripotent stem cell-derived neural precursor cells （hiPSC-NPCs） on decellularized optic nerve （DON）， so as to provide a promising approach for repair of nerve tissue injury. MethodsThrough directional induction and tissue engineering technology， human induced pluripotent stem cells （hiPSCs） and 3D DON scaffolds were combined to construct neural network-like tissues. Then the hiPSCs were directionally induced into human neural precursor cells （hNPCs） and neurons. Immunofluorescence staining was used to identify cell differentiation efficiency. 3D DON scaffolds were prepared. Morphology and cytocompatibility of scaffolds were identified by scanning electron microscopy and Tunnel staining. Induced hiPSC-NPCs were seeded on DON scaffolds. Immunofluorescence staining， scanning electron microscopy， transmission electron microscopy and patch clamp were used to observe the morphology and functional identification of constructed neural network tissues. Results①The results of immunofluorescence staining suggested that most of hiPSC-NPCs differentiated into neurons in vitro. We had successfully constructed a neural network dominated by neurons. ② The results of scanning electron microscopy and immunohistochemistry suggested that a neural network-like tissue with predominating excitatory neurons in vitro was successfully constructed. ③The results of immunohistochemical staining， transmission electron microscopy and patch clamp indicated that the neural network-like tissue had synaptic transmission function. ConclusionA neural network-like tissue mainly composed of excitatory neurons has been constructed by the combination of natural uniform-channel DON scaffold and hiPSC-NPCs， which has the function of synaptic transmission. This neural network plays a significant role in stem cell derived replacement therapy， and offers a promising prospect for repair of spinal cord injury （SCI） and other neural tissue injuries.
