大鼠脊髓片器官培养模型的建立

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。

The establishment of the model of the organotypic culture from the SD rats'''' spinal cord

Objective: To establish a model of spinal cord organotypic culture in vitroof Sprague Dawlay (SD) rat. Methods The lumber spinal cords prepared from the 6-day -old mice were cut into 350 μm coronary and then transferred onto Millicell-CM cultured plate inserts and grew . They were divided into 8 groups according the culturing time and assessed the viability of the culture by the tetrazolium dye (MTT) , observe the cytoarchitecture by cresyl violet staining and count the motor neurons by cholineacetyl transferase (ChAT) immunohistochemical staining. Results:With the elongation of the culture time, although the viability and the number of the motor neurons of the cultured spinal cord decreased, the cytoarchitecture was kept. Conclusion:This organotypic culture of the spinal cord from mice is simple and effective method for cultured model in vitro.