中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (2): 339-345.doi: 10.4103/1673-5374.244797

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

原代培养来源于大鼠背根神经节高纯度卫星胶质细胞的新方法

  

  • 出版日期:2019-02-15 发布日期:2019-02-15
  • 基金资助:

    国家自然科学基金(31560295); 云南省科技厅优先联盟基金会和中国昆明医科大学基金(2015FB098); 中国昆明医科大学重大科技成果培育项目(CGPY201802); 中国云南省卫生科技计划项目(2014NS202)

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

Xian-Bin Wang 1 , Wei Ma 1, Tao Luo 1, 2 , Jin-Wei Yang 1, 3 , Xiang-Peng Wang 1 , Yun-Fei Dai 1 , Jian-Hui Guo 3 , Li-Yan Li 1   

  1. 1 Institute of Neuroscience, Kunming Medical University, Kunming, Yunnan Province, China
    2 Medical Faculty, Panzhihua University, Panzhihua, Sichuan Province, China
    3 Second Department of General Surgery, First People’s Hospital of Yunnan Province, Kunming, Yunnan Province, China
  • Online:2019-02-15 Published:2019-02-15
  • Contact: Li-Yan Li, MD, kmliyanl@163.com; Jian-Hui Guo, guojianhuikm@163.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 31560295; the Priority Union Foundation of Yunnan Department of Science and Technology and Kunming Medical University of China, No. 2015FB098; the Project of Major Scientific and Technological Achievements Cultivation of Kunming Medical University of China, No. CGPY201802; the Health Science and Technology Plan Projects of Yunnan Province of China, No. 2014NS202.

摘要:

以往对大鼠背根神经节卫星胶质细胞的原代培养方法尚不十分成熟;为进一步完善该类细胞的原代培养方法,实验收集新生大鼠背根神经节,并在潮湿的5%CO2培养箱(37℃)中培养。(1)培养3d后,一些细胞从背根神经节迁移;(2)传代培养后,免疫荧光染色法显示培养的细胞表达3种卫星胶质细胞特异性标记物:谷氨酰胺合成酶,胶质原纤维酸性蛋白(GFAP)和S100β,表明培养的细胞为卫星胶质细胞,且纯度> 95%;(3)实验成功地建立了一种新的原代培养方法,可以获得高纯度的大鼠背根神经节的卫星胶质细胞。

orcid: 0000-0001-7148-4901 (Jian-Hui Guo)

关键词: 细胞培养, 背根神经节, 免疫荧光鉴定, 卫星神经胶质细胞, 神经再生

Abstract:

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.

Key words: nerve regeneration, cell culture, dorsal root ganglia, immunofluorescence identification, satellite glial cells, neural regeneration