视网膜细胞培养上清液对胚胎干细胞定向分化的神经样细胞钠通道动力学的影响

Effect of supernatant fluid of retinal cells on dynamics of sodium ion channels of neuron-like cells derived from embryonic stem cells

目的探讨不同视网膜细胞上清液对由胚胎干细胞诱导的神经细胞生理机能的影响。方法对拟胚体分别使用视黄酸（A组）、视黄酸＋鼠视网膜胶质细胞和神经细胞培养上清液（B组）、视黄酸＋人胎儿视网膜胶质细胞培养上清液（C组）进行次级诱导。在诱导后5～21d，对各组中的细胞分别行细胞膜上的钠离子通道分析。结果诱导后5～21d时，各组细胞钠电流变化均不明显。A组细胞钠通道呈爆发状的放电现象；B组呈现短促而频繁的状态；C组开放时程较长。三组细胞中，无电流细胞的比例分别为25%、11.4%和23.8%，但组间差异无统计学意义（P〉0.05）。A组Na^＋电流细胞数目表现为先升高后下降，B组表现为一直上升，C组表现为先下降后趋于平稳。A组细胞通道的开放时间最长，B组最短。三组的开放时间分布均可使用双指数拟和。结论在胚胎干细胞向神经细胞诱导中，视网膜细胞上清液对诱导细胞的生理功能会产生明显影响。

Objective To study the effect of different types of supernatants fluid of retinal cells on the physiological function of neuron cells derived from embryonic stem cells. Methods Embryonic bodies were sub-induced by retinoic acid （group A）, retinoic acid with the supernatant fluid of retinal glia cells and neurons of mouse （group B）, retinoie acid with the supernatant fluid of fetal retinal glia cells （group C）, respectively, The Sodium ion channels on the cytomembrane in the 3 groups were analyzed 5-21 days after the inducement. Results The sodium current in each group didn＇t change much 5 21 days after the inducement. The sodium channels presented burst-opening discharge in group A, brief-opening discharge in group B, and long-opening discharge in group C. The percentage of the cells without current in group A, B and C was 25%, 11.4%, and 23.8%, respectively, but the difference was not significant among the 3 groups（P〉0. 05）. The number of cells with sodium current increased at first and decreased later in group A, continuously increased in group B, and decreased at first and kept stable later in group C. The open time of sodium channels was the longest in group A, and the shortest in group B. The distribution of open time in the three groups could be managed with two-step exponential fit. Conclusion The supernatant fluid of retinal cells has apparent influence on the physiological function of the neuron cells derived from embryonic stem cells.