GDNF及不同生长底物对培养颈上神经节交感神经元的作用

目的 探讨胶质细胞源性神经营养因子及不同生长底物对培养颈上神经节交感神经元存活和突起生长的影响.方法 取新生大鼠颈上神经节(superior cervical ganglion, SCG)体外分离培养,培养基中含不同浓度的胶质细胞源性神经营养因子(glial cell derived neurotrophic factor, GDNF),并选取不同方法溶解多聚-D-赖氨酸(poly-D-lysine, PDL)包被培养盖片,观测交感神经元的存活和突起生长状况.结果 用0.01 mol/L硼酸溶解的PDL进行包被,细胞存活和突起生长最好,且神经元分散好,优于其他各组;GDNF对培养交感神经元的存活、突起生长具有显著的促进作用,且与其浓度存在量效关系,但它并不能阻止培养前5 d存活神经元数量的减少.结论 用0.01 mol/L硼酸溶解PDL包被是一种良好的生长底物处理方法,GDNF能促进离体培养的交感神经元的存活和突起生长.

Effects of GDNF and substrate on superior cervical ganglion sympathetic neurons in vitro

Objective To explore the effects of glial cell-derived neurotrophic factor and different substrates on cell survival and neurite growth of superior cervical ganglion sympathetic neurons from neonatal rat in vitro. Methods Sympathetic neurons from superior cervical ganglion (SCG) of neonatal rat respectively were dissociated and cultured. The glial cell-derived neurotrophic factor (GDNF) of different quantities were added in the medium, and culture cover slips were coated by variant methods with poly-D-lysine (PDL). The cell survival and neurite growth of cultured sympathetic neurons were observed by phase-contrast microscopy. Results In the group of cover glasses coated with PDL dissolved by 0.01 mol/L boric acid solution, cells grew dispersedly well, and both the cell survival and neurite growth were better than others. GDNF promoted the cell survival and neurite growth of sympathetic neurons markedly, and there was a dose-effect relationship, but it could not abolish the decrease of survival cells from 1 d to 5 d in vitro. Conclusion It is a favorable substrate that coated with PDL dissolved 0.01 mol/L by boric acid solution, and both the cell survival and neurite growth of sympathetic neurons can be improved by GDNF in vitro.