摘要
目的探讨离体培养脊髓片的理想厚度和大鼠的最佳周龄。方法通过不同周龄的大鼠(1周、2周、3周、4周龄各10只,共40只),以及不同厚度(切片厚度分为:100μm、200μm、300μm以及400μm 4种)的脊髓组织切片培养,采用MTT技术进行脊髓片培养细胞的活性鉴定,观察脊髓片培养不同时间点其神经细胞的活性。结果1~7 d鼠龄的脊髓组织片培养1~14 d时细胞活性最高,但其神经细胞结构形态容易受到破坏,不利于观察,且在培养超过14 d时神经细胞活性明显下降;2~3周鼠龄的脊髓组织片培养细胞,在培养14 d后仍有较高的神经细胞的活性并保持较好的细胞形态结构;1~4各周龄组大鼠200μm和300μm左右厚度的脊髓片培养神经细胞活性最稳定(85%~95%)。结论进行离体脊髓片培养,选择7~14 d(即1~2周龄)大鼠作为取材对象较为理想,选择200μm^300μm切片厚度的脊髓片进行培养较为合适。
Objective To explore the ideal thickness of spinal cord slices and the optimal age of Sprague-Dawley rat for culturing in vitro. Methods Forty Sprague-Dawley rats aged from 1-4 weeks were randomly divided into four groups （ every group, n = 10）, and the thickness of cultured spinal cord slices included 100μm, 2μm, 300μm and 400μm respectively in four groups. The activity of the culture slices were evaluated by MTT technique and observed at different times. Results The activity of spinal cord slices from the rats aged 1-7 days achieved the highest when cultured from 1-14 days, but the structure were apt to be destroyed, and the activity decreased rapidly when the culture time exceeded 14 days. The spinal cord slices from the rats aged from 2-3 weeks still had a higher activity and maintained a better structure after cultured over 14 days. The activity of spinal cord slices with the thickness of 200μm and 300μm were the best in each age group. Conclusion The activity of spinal cord slices cultured in vitro can be affected by the slice thickness, culture time and the age of rats. The thickheSS of 200μm and 300μm and the age of 1-2 weeks are the ideal conditions for the activity of spinal cord slices cultured in vitro.
出处
《脊柱外科杂志》
2006年第2期121-124,共4页
Journal of Spinal Surgery
基金
国家自然科学基金(39900148)
关键词
脊髓
神经元
细胞培养
大鼠
spinal cord
neurons
cell culture
rats
