大鼠嗅粘膜原代培养细胞的分类及细胞社会学特性研究

目的：探讨嗅粘膜的细胞社会学特性，为嗅粘膜混合细胞移植治疗中枢神经系统损伤提供可行性研究资料。方法：自成年大鼠鼻中隔活体剪取部分嗅粘膜制成细胞悬液接种于玻璃培养皿，用含10％胎牛血清的F12培养基培养，动态观察细胞的贴壁、生长及分化过程，并用波形蛋白(vimentin)、层粘连蛋白(Iaminin)、巢蛋白(nestin)、神经元特异性烯醇化酶(neuron specific enolase，NSE)、胶质纤维酸性蛋白(glial fibrillary acidic protein，GFAP)及神经生长因子受体p^75(nerve growth factor receptor p^75，NGFRp^75)抗体作免疫细胞化学染色，对阳性细胞进行形态学鉴定。结果：体外培养的嗅粘膜细胞中包含成纤维细胞、神经干细胞、嗅细胞和嗅鞘细胞等，不同类型细胞的形态及生长特性存在差异。结论：嗅粘膜细胞之间具有协调的细胞社会学关系；成体嗅粘膜混合细胞群移植治疗中枢神经系统损伤的可行性具有细胞社会学基础。

Cytosociological Characteristics and Classification of Cultured Cells from Nasal Olfactory Mucosa of Adult Rat

Objective:To explore applicability of transplanting cultured cells of nasal olfactory mucosa to repair the injured central nervous system, the cytosociological characteristics and classification of the cells from adult rat were studied. Methods:The nasal olfactory mucous membrane was alive removed from the adult SD rats, the cells of the mucous membrane were culture in F12 medium contained 10% fetal bovin serum. The growth and morphologic characteristics of cultured cells were observed by immunocytochemistry using antibodies for nestin, neuron specific enolase (NSE), glial fibrillary acidic protein(GFAP), nerve growth factor receptor p 75(NGFRp 75), vimentin and laminin. Results:The group of cultured cells from nasal olfactory mucosa of adult rat containd neural stem cells, fibroblasts, olfactory neurons, olfactory ensheathing cells ect. There was difference in morphology and cytobiology among the different cell types. The cells attached to the bottom of culture vessels first were flat and immunopositive for vimentin and laminin. The spherecal nestin positive cells were adhered to the flat substrate cells then formed clonal spheres by mitosis. The clonal spheres were positive for nestin as well. GFAP and NGFRp 75 immunopositive cells were spherecal, astroid or spindle-shaped,which were also positive for laminin. A few cells were positive for NSE. Conclusion:There was cooperative cytosociological relationship in the cell group; Transplanting the cell group to repair the injured central nervous system is applicable in cytosociology.