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目的通过不同培养时间细胞的存活率和形态学特征了解体外培养的脊髓神经元的生长规律,以便选择施加干预因素及观测结果的最佳时间。方法在倒置显微镜(×250)和高倍镜视野(×400)下,利用图像分析系统测量培养1 d、3 d5、d7、d、9 d CHAT阳性细胞的轴突长度、树突数目、树突总长度、树突分叉点数目和胞体面积5个形态学指标。结果存活率在第3~5 d维持在50%左右。细胞突起长度、突起数目、胞体面积前3 d增长幅度最快,第4~7 d各项指标缓慢上升,生长稳定,7 d后各项指标快速下降。结论原代培养的脊髓运动神经元的存活率、突起长度、分叉点数目和胞体面积等指标可以反映细胞的生长状况,适宜于实验中各项指标的观测。
OBJECTIVE: To understand the growth pattern of neurons cultured in vitro by the survival rate and morphological characteristics of cells at different culture periods in order to choose the optimal time for intervention and observation. Methods The length of axon, the number of dendrites, the number of dendrites, the number of dendrites in CHAT positive cells cultured on d 1, d 3, d 7, d 9 and d 9 were measured under an inverted microscope (× 250) and high magnification field (× 400) Total length, the number of dendritic bifurcation points and cell body area of 5 morphological indicators. Results The survival rate maintained at about 50% on the 3rd to 5th days. The number of protuberances, the number of protuberances, and the area of the somatic cells increased most rapidly in the first 3 days. The indexes increased slowly and steadily on the 4th to 7th days, and all the indexes decreased rapidly after 7 days. Conclusion The survival rate, the length of protuberance, the number of bifurcation points and the cell body area of primary cultured spinal motor neurons can reflect the growth of cells and is suitable for the observation of various indexes in the experiment.