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目的:通过器官培养的方法建立大鼠器官型脊髓片培养模型。方法:取出生后6天的Sprague Dawlay(SD)乳鼠的腰段脊髓作冠状切片后将脊髓片置于Millicell～CM膜上使脊髓处于液气交界面上进行培养,取培养不同时间点的脊髓片,采用倒置显微镜、胆碱乙酰转移酶免疫组化及焦油紫(Nissle)染色、四甲基偶氮唑盐(MTT)测细胞活性等方法进行观察。结果:在15天之内,虽然随着培养时间的延长,脊髓片组织内的运动神经元数目及细胞活性检测结果呈递减趋势,但运动神经元的形态结构保持完好,细胞层次仍然保留。结论:利用微孔膜进行器官型脊髓片培养是一种简便有效的神经组织体外培养方法。
OBJECTIVE: To establish a rat organ-type spinal cord slice culture model through organ culture. Methods: The spinal cord of Sprague Dawley (SD) suckling rat was coronal sectioned 6 days after birth. The spinal cord was placed on Millicell ~ CM membrane and the spinal cord was placed on the liquid-gas interface. The cultured cells were cultured at different time points The spinal cord slices were observed by inverted microscope, immunohistochemistry of choline acetyltransferase, Nissle staining and MTT assay. Results: Within 15 days, although the number of motor neurons and the activity of cell activity in spinal cord tissue decreased with the prolongation of culture time, the morphological structure of motor neurons remained intact and the cell level remained. Conclusion: The use of microporous membrane for organotypic spinal cord culture is a simple and effective method of nerve tissue culture in vitro.