A novel primary culture method for high-purity satellite glial cells derived from rat dorsal root ganglion

论文摘要

Satellite glial cells surround neurons within dorsal root ganglia. Previous studies have focused on single-cell suspensions of cultured neurons derived from rat dorsal root ganglia. At present, the primary culture method for satellite glial cells derived from rat dorsal root ganglia requires no digestion skill. Hence, the aim of the present study was to establish a novel primary culture method for satellite glial cells derived from dorsal root ganglia. Neonatal rat spine was collected and an incision made to expose the transverse protrusion and remove dorsal root ganglia. Dorsal root ganglia were freed from nerve fibers, connective tissue, and capsule membranes, then rinsed and transferred to 6-well plates, and cultured in a humidified 5% CO2 incubator at 37°C. After 3 days in culture, some cells had migrated from dorsal root ganglia. After subculture, cells were identified by immunofluorescence labeling for three satellite glial cell-specific markers: glutamine synthetase, glial fibrillary acidic protein, and S100β. Cultured cells expressed glutamine synthetase, glial fibrillary acidic protein, and S100β, suggesting they are satellite glial cells with a purity of > 95%. Thus, we have successfully established a novel primary culture method for obtaining high-purity satellite glial cells from rat dorsal root ganglia without digestion.

论文目录

• Graphical Abstract
• Introduction
• Materials and Methods
•   Study approval and animals
•   DRG preparation and primary culture
•   Cell subculture and purification
•   Immunofluorescence
•   Immunohistochemistry on SGC in DRG
• Results
•   Morphological characteristics of primary cultured SGCs
•   Morphological characteristics of SGCs after subculture
•   Characterization of cultured SGCs by immunofluorescence
• Discussion
•   Recent developments in DRG-SGC culture systems and their applications
•   Factors affecting survival and proliferation of SGCs in vitro
•   Purification and culture of SGCs
• Conclusion

文章来源

类型: 期刊论文

作者: Xian-Bin Wang,Wei Ma,Tao Luo,Jin-Wei Yang,Xiang-Peng Wang,Yun-Fei Dai,Jian-Hui Guo,Li-Yan Li

来源: Neural Regeneration Research 2019年02期

年度: 2019

分类: 医药卫生科技,基础科学

专业: 生物学,基础医学

单位: Institute of Neuroscience, Kunming Medical University,Medical Faculty, Panzhihua University,Second Department of General Surgery, First People's Hospital of Yunnan Province

基金: supported by the National Natural Science Foundation of China,No.31560295(to LYL),the Priority Union Foundation of Yunnan Department of Science and Technology and Kunming Medical University of China,No.2015FB098(to JHG),the Project of Major Scientific and Technological Achievements Cultivation of Kunming Medical University of China,No.CGPY201802(to LYL),the Health Science and Technology Plan Projects of Yunnan Province of China,No.2014NS202(to JHG)

分类号: R329.2

页码: 339-345

总页数: 7

文件大小: 532K

下载量: 54