CAJ | 학술논문

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70 000～150 000及相对分子质量150 000～300 000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-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-多聚赖氨酸包被的孔内细胞少量存活;相对分子质量70 000～150 000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150 000～300 000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150 000～300 000的L-多聚赖氨酸最适宜神经元的生长.
목적 탐토불동상대분자질량적좌선(L-)다취뢰안산대신생서원대해마신경원생장적영향. 방법 채용침예제압과망법제비유리신생서해마신경세포현액,장세포분위3조,분별충식우미포피、상대분자질량70 000～150 000급상대분자질량150 000～300 000적L-다취뢰안산포피적배양판내.Neurobasal-A선택성배양기유지배양,관찰불동계단신경원적형태,신경원특이성희순화매면역형광염색감정병계산순도,관찰불동상대분자질량적L-다취뢰안산대신경원생장특성적영향. 결과 신경원충식1h후개시첩벽；4d후신경원포체투량,결구완정,돌촉장출2～3지；8d후포체성숙,돌촉증다,교직성망.신경원특이성희순화매면역형광염색감정위양성,신경원평균순도위92.6％+4.62％.무L-다취뢰안산포피적공내세포소량존활;상대분자질량70 000～150 000적L-다취뢰안산포피적공내세포포단양생장;상대분자질량150 000～300 000적L-다취뢰안산포피적공내세포생장성균균적단층,교소유포단현상. 결론 불동상대분자질량적L-다취뢰안산영향신경원적첩부화행위,상대분자질량150 000～300 000적L-다취뢰안산최괄의신경원적생장.
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.