生长底物对培养胚胎大鼠脊髓运动神经元的作用

Effect of different substrates coated on spinal motor neurons from embryonic rat cultured in vitro

目的 :为了研究几种生长底物对培养的胚胎大鼠脊髓运动神经元生长特性的作用。方法 :采用胚胎大鼠脊髓腹侧组织进行分离培养 ,建立脊髓运动神经元原代分散细胞培养 ,并应用免疫组织化学染色进行鉴定 ;选取生长底物如多聚赖氨酸 (PLL)、Ⅰ型胶原、层粘连蛋白 (LN)、PLL和LN联合等进行包被 ,观察脊髓运动神经元的生长特性。结果 :PLL用 0 0 1mol L硼酸溶解 ,细胞生长良好 ;PLL和LN联合包被时 ,脊髓运动神经元存活率高 ,细胞分散良好。结论 :PLL和LN联合包被培养脊髓运动神经元 。

Objective:To study the effect of different substrates coated on the cell survival and neurite outgrowth of spinal motor neurons (SMN) from embryonic rat cultured in vitro.Methods:The ventral spinal tissue was isolated from embryonic rats and digested into dissociated cell suspention for culture,then the cells were identified as SMN by immunohistochemistry stain.Poly-L-lysine(PLL) was dissolved into distilled water,phosphate-buffered saline solution,boric acid at 0.1 and 0.01 mol/L concentration respectively.The different substrates include PLL,collagen Ⅰ,laminin and PLL combined with laminin.Distilled water was used as control.The neuron survival numbers and the mean length of the neurites were measured and compared.Results:The cells on the PLL dissolved into boric acid at 0.01 mol/L concentration survived well.The SMNs grown on the PLL combined with laminin were in dispersed disitribution with high survival rate.Conclusion:PLL combined with laminin is the best for the study of the motor neuron including both soma and neurite.