Characteristics of neural growth and cryopreservation of the dorsal root ganglion using three-dimensional collagen hydrogel culture versus conventional culture  

作　　者：Ze-Kai Cui Shen-Yang Li Kai Liao Zhi-Jie Wang Yong-Long Guo Luo-Sheng Tang Shi-Bo Tang Jacey Hongjie Ma Jian-Su Chen 

机构地区：[1]Department of Ophthalmology,the Second Xiangya Hospital,Central South University,Changsha,Hunan Province,China [2]Aier Eye Institute,Changsha,Hunan Province,China [3]Aier School of Ophthalmology,Central South University,Changsha,Hunan Province,China [4]Institute of Ophthalmology,Medical College,Jinan University,Guangzhou,Guangdong Province,China [5]Department of Ophthalmology,First Affiliated Hospital of Jinan University,Guangzhou,Guangdong Province,China [6]Key Laboratory for Regenerative Medicine of Ministry of Education,Jinan University,Guangzhou,Guangdong Province,China [7]CAS Center for Excellence in Brain Science and Intelligence Technology,Chinese Academy of Sciences,Shanghai,China [8]Imaging and Functional Center,Guangzhou Aier Eye Hospital,Guangzhou,Guangdong Province,China

出　　处：《Neural Regeneration Research》2021年第9期1856-1864,共9页中国神经再生研究（英文版）

基　　金：supported by the National Natural Science Foundation of China,Nos.82000871(to ZKC),81871495(to JSC);the Natural Science Foundation of Hunan Province,China,No.2020JJ5001(to ZKC);the Science Research Grant of Aier Eye Hospital Group,China,No.AF1913D2(to ZKC);Central South University Postdoctoral Funds,China。

摘　　要：In vertebrates, most somatosensory pathways begin with the activation of dorsal root ganglion(DRG) neurons. The development of an appropriate DRG culture method is a prerequisite for establishing in vitro peripheral nerve disease models and for screening therapeutic drugs. In this study, we compared the changes in morphology, molecular biology, and transcriptomics of chicken embryo DRG cultured on tissue culture plates(T-DRG) versus three-dimensional collagen hydrogels(C-DRG). Our results showed that after 7 days of culture, the transcriptomics of T-DRG and C-DRG were quite different. The upregulated genes in C-DRG were mainly related to neurogenesis, axon guidance, and synaptic plasticity, whereas the downregulated genes in C-DRG were mainly related to cell proliferation and cell division. In addition, the genes related to cycles/pathways such as the synaptic vesicle cycle, cyclic adenosine monophosphate signaling pathway, and calcium signaling pathway were activated, while those related to cell-cycle pathways were downregulated. Furthermore, neurogenesis-and myelination-related genes were highly expressed in C-DRG, while epithelial–mesenchymal transition-, apoptosis-, and cell division-related genes were suppressed. Morphological results indicated that the numbers of branches, junctions, and end-point voxels per C-DRG were significantly greater than those per T-DRG. Furthermore, cells were scattered in T-DRG and more concentrated in C-DRG, with a higher ratio of 5-ethynyl-2′-deoxyuridine(EdU)-positive cells in T-DRG compared with C-DRG. C-DRG also had higher S100 calcium-binding protein B(S100 B) and lower α-smooth muscle actin(α-SMA) expression than T-DRG, and contained fewer terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)-positive cells after 48 hours of serum starvation. After cryopreservation, C-DRG maintained more intact morphological characteristics, and had higher viability and less TUNEL-positive cells than T-DRG. Furthermore, newly formed nerve bundles were able to grow along

关 键 词：ANTI-APOPTOSIS collagen hydrogel CRYOPRESERVATION dorsal root ganglion NEUROGENESIS RNA-seq Schwann cell tissue engineering 

分 类 号：R338[医药卫生—人体生理学]