大鼠小团样视网膜神经细胞培养的方法

Method of rat retinal neural cell culture

目的:探讨视网膜神经细胞的体外培养方法。方法:取出生3d的SD大鼠视网膜组织,木瓜蛋白酶和胰蛋白酶依次消化并吹打制成含不同大小细胞团块的细胞悬液,放入50mL/LCO2,37℃的孵箱中培养。用倒置相差显微镜观察视网膜神经细胞的生长情况。结果:聚集成小团的视网膜神经细胞(<50个细胞)与完全分离成单个的神经细胞相比,贴壁和发出轴突的时间早,轴突长度长,细胞间的轴突联系增多,细胞存活时间亦延长。而较大团块状的细胞只可以见到轴突的伸出和伸长。结论:小团样的视网膜神经细胞培养有利于神经细胞的存活和细胞间的轴突联系,这可能得益于视网膜多种细胞间的联系在体外的持续保持。

AIM： To explore culture methods in vitro of rat retinal neural cell culture. METHODS： Neural retina of postnatal 3 days Sprague-Daw rats were digested by papain and trypsin in turn, and then gently percussed to loose adherent cells. Cell suspension with different sizes of cell aggregates were maintained at 37℃ in a humidified 50mL/L CO2 incubator. Cell growth was observed by phase-contrast microscope. RESULTS： Compared with single dissociated cell, cells in small cell aggregates had more axonal connection and longer survival time, while cells in big cell aggregates only showed axonal growth. CONCLUSION： Retinal neural cell culture is conducive to survival of nerve cells and cell contact between the axon, which may due to the continual remaining of intercellular connection in vitro.