摘要
目的探讨不同相对分子质量的左旋(L-)多聚赖氨酸对新生鼠原代海马神经元生长的影响。方法采用针蕊挤压过网法制备游离新生鼠海马神经细胞悬液,将细胞分为3组,分别种植于未包被、相对分子质量70000~150000及相对分子质量150000~300000的L.多聚赖氨酸包被的培养板内。Neurobasal。A选择性培养基维持培养,观察不同阶段神经元的形态,神经元特异性烯醇化酶免疫荧光染色鉴定并计算纯度,观察不同相对分子质量的L一多聚赖氨酸对神经元生长特性的影响。结果神经元种植1h后开始贴壁:4d后神经元胞体透亮.结构完整,突触长出2~3支;8d后胞体成熟,突触增多,交织成网。神经元特异性烯醇化酶免疫荧光染色鉴定为阳性,神经元平均纯度为92.6%±4.62%。无L-多聚赖氨酸包被的孑L内细胞少量存活:相对分子质量70000~150000的L-多聚赖氨酸包被的孔内细胞抱团样生长;相对分子质量150000~300000的L.多聚赖氨酸包被的孔内细胞生长成均匀的单层,较少有抱团现象。结论不同相对分子质量的L-多聚赖氨酸影响神经元的贴附和行为,相对分子质量150000~300000的L-多聚赖氨酸最适宜神经元的生长。
Objective To investigate the effects of poly-L-lysine of different molecular weights on the growth of primary hippocampal neurons in neonatal rats. Methods The hippocampal neurons from neonatal rats were prepared with the method of extrusion by needle core; and then, the hippocampal neurons were divided into three groups and respectively planted in culture plates where coated poly-L-lysine of different molecular weights (no poly-L-lysine, 70 000-150 000 and 150 000- 300 000). The neurons were maintained in Neurobasal-A medium without fetal bovine serum. The neurons were viewed at different time points and indentified by neuron specificity enolization enzyme immunofluorescence staining; the purity of the cells was calculated and the effects of poly-L-Lysine of different molecular weights on the growth of cells were observed. Results The neurons attached to the culture plates 1 h after the plantation. Four d after the plantation, the neurons had shining body, integrity stucture, having 2-3 synapses. Eight d after the plantation, the neurons became mature; the axons of neurons interweaved into the net; the cells were identified as neurons with an average purity of (92.6+ 4.62)%. All neurons without poly-L-lysine almost died; the cells in the plates of poly-L-lysine of 70 000-150 000 distributed uniformly. Conclusion Different molecular weights of poly-L-lysine can affect the neurons adhesion and behavior; poly-L-lysine with large molecular weight (150 000-300 000) is most suitable for neurons.
出处
《中华神经医学杂志》
CAS
CSCD
北大核心
2013年第7期649-652,共4页
Chinese Journal of Neuromedicine
基金
国家自然科学基金(81100597)
重庆市自然科学基金(2009BB5323)