大鼠视网膜神经元的原代培养方法

目的:探求视网膜神经元的最佳分离和培养方法,提高原代培养神经元的纯度及活性。方法:采用新生大鼠视网膜神经元,应用含100mL/L新生牛血清、100g/LF-12 Nutrient Mixtures的DMEM培养基进行接种培养,含20g/LB-27 Serum-Free Supplements的Neurobasal Medium进行维持培养,利用尼氏染色进行神经元鉴定。结果:培养的神经元生长良好,胞体饱满,突起长。尼氏染色示神经元比例大于90%。结论:采用机械分离法和Neurobasal Medium培养基可以使新生大鼠视网膜神经元得到良好的生长和较高的纯度。

Primary culture of retinal neurons of newborn rat

·AIM:To explore the best method of separating and culturing retinal neurons,and to improve purity and activity of primary cultured neurons..METHODS:Primary culture of retinal neurons of newborn rat was performed by Dulbecco’s modified Eagle’s medium(DMEM)with 100mL/L newborn calf serum and 100g/L F-12 Nutrient Mixtures.The Neurobasal Medium with 20g/L B-27 Serum-Free Supplements was used for maintenance culture.Evaluation of neurons was made by Nissl’s staining..RESULTS:Cultured neurons grew well.They were plump with long processes.The purity of neurons was higher than 90%..CONCLUSION:Primary culture of retinal neurons of new born rat by mechanical separation and Neurobasal Medium is a reliable way to obtain active neurons with high purity..