基质细胞衍生因子对海马神经细胞迁移的影响

Stromal derived factor 1 alpha alters migration of hippocampal cells cultured in vitro

目的观察不同浓度的SDF1alpha对体外培养的海马神经细胞迁移的影响。方法通过Transwell小室方法观察细胞在轴突导向因子诱导下穿过多孔滤膜的情况。将分离的海马神经细胞种入上室内,并且上室内细胞培养基均为无血清培养基(饥饿疗法),下室内则分别采用含有0、50、100、150、200、250、300ng/ml的SDF1alpha的培养基(对照组含有0ng/ml的SDF1alpha)12h后,观察迁移的神经细胞数量,统计后找到最佳实验剂量。采用最佳实验剂量作用细胞24h,观察神经细胞形态学的变化。结果不同浓度的SDF1alpha作用12h后,迁移的神经细胞数量分别为16.67±1.21、16.83±2.14、35.50±4.32、36.33±1.21、36.83±1.94和38.17±2.71,与对照组迁移的神经细胞数量9.17±2.14相比,有显著性差异。采用150ng/ml的SDF1alpha作用细胞24h,神经细胞突起延长。结论 SDF1alpha可以促进体外培养的海马神经细胞迁移和突起延长。

Objective Stromal cell-derived factor-1（SDF1） alpha,a kind of chemotactic protein,is secreted by marrow stromal cells to exert its function in immune system.Recently,studies have shown that SDF1alpha expresses in the brain,which suggests that SDF1alpha may play an important role in nervous system.This study is to culture hippocampal cells in vitro to detect the function of SDF1alpha on cell migration and neurite outgrowth.Methods Primary hippocampus neurons were obtained from newborn SD rats.Bilateral hippocampi were carefully dissected and digested in 0.25% trypsin at 37℃ for 5 minutes.The in vitro migration study was carried out with transwell assay.Cells were plated into the upper layer of the transwell device.Medium with varying doses of SDF1alpha were placed into the lower layer.After 12-hour incubation at 37℃ in 5% CO2,the incubation medium was discarded and the membrane was washed with PBS twice.The cells on the upper layer of the filtering film were wiped away with cotton.The filtering film was carefully cut out and fixed by methanol for 2h at room temperature.The filtered cells were counted and calculated.Results Migration of hippocampal cells induced by different doses of SDF1alpha were 16.67±1.21,16.83±2.14,35.50±4.32,36.33±1.21,36.83±1.94 and 38.17±2.71 respectively.And cell migration of control group was 9.17±2.14.There were significant differences between control and experiment group.When 150ng/ml SDF1 was added into the culture system,neurite outgrowth was observed in the culture hippocampal cells.Conclusions SDF1alpha may alter migration and neurite outgrowth of hippocampal cells cultured in vitro.