大鼠运动区脑片的培养方法

Rat Motor-area Brain Slice Culture

利用出生后1天的乳鼠大脑活组织切片建立脑片的培养模型,并用抗非磷酸化神经丝单克隆抗体SMI-32对皮层锥体细胞加以鉴定、计数,与生长至同时期的大鼠运动区皮层锥体细胞数目比较,并测定不同时期培养液中乳酸脱氢酶(LDH)含量。结果显示脑片体外生长良好,形态完整,皮层锥体细胞数目恒定,与体内生长的锥体细胞数目无显著差异,培养液中LDH含量稳定。大鼠脑片的培养以及锥体细胞组化鉴定技术为研究有关皮层运动神经元的疾病如肌萎缩侧索硬化提供了有效的方法。

Brain slice cultures were prepared using brains from 1-day-old rat pups. Pyramidal cells＇ survival was evaluated by anti-nonphosphorylated neurofilament H （SMI-32） immunohistochemistry staining. Compared the pyramidal cells with those in vivo, lactate dehydrogenase （LDH） level in culture mediums were also measured. The brain explants could be maintained in culture for more than 1 month with excellent form and a stable population of cortical pyramidal cells in layer V. The level of LDH at different culture times had no significant differences. Rat brain slice cultures and identification of pyramidal cells provided an effective method to Study the diseases involved in the cortical motor neuron.