成年大鼠嗅粘膜细胞原代培养及形态学研究

PRIMARY CULTURE AND MORPHOLOGICAL STUDY OF OLFACTORY ENSHEATHING CELLS FROM THE ADULT RAT OLFACTORY MUCOSA

为了探讨大鼠嗅粘膜嗅鞘细胞(OECs)的分离与纯化培养方法并对其形态学进行研究,我们自鼻腔嗅粘膜取材后采用差速贴壁、机械刮除和阿糖胞苷(Arac)抑制相结合的纯化培养方法培养嗅粘膜细胞,并分不同阶段进行镜下观察及p75NGFR和GFAP免疫组化染色鉴定。结果显示,自嗅粘膜组织可以培养出四种细胞:梭形细胞、双极细胞、窝形细胞和大细胞。其中大部分梭形和双极细胞呈p75NGFR或GFAP免疫反应阳性,属于OECs。结果提示本文介绍的培养方法可以培养出嗅粘膜OECs;差速贴壁、机械刮除和Arac抑制相结合方法可纯化OECs。

In order to find an effective method to puriy olfactory ensheathing cells (OECs) from the olfactory mucosa of adult rat and to investigate the morphological changes of the cultured OECs, the nasal olfactory mucosa was harvested from the posterior part of the nasal septum and cultured, purified with the method of combining the different rates of attachment among the various cell types, mechanical curettage and the Ara-c inhibition. The morphological changes of cultured OECs were observed at different stages and identified with both nerve growth factor receptor p75 (NGFRp75) and glial fibrillar acidic protein (GFAP) immunohistochemistry. The results showed that the cultured mucosa cells presented mainly four morphological types: fusiform cell, bipolar cell, nest cell and giant cell. Part of these bipolar or fusiform cells are immunohistochemically positive for NGFRp75 and GFAP, indicating that they belong to OECs. These results suggest that OECs were able to be cultured and purified through combining the different rates of attachment among the various cells types, mechanical curettage and the Ara-c inhibition.