年轻成年GFP大鼠嗅鞘细胞的分离培养与形态学特征

目的 建立年轻成年GFP大鼠嗅鞘细胞(OECs)的分离培养方法 ,为OECs移植治疗脊髓损伤奠定基础. 方法 解剖显微镜下、无菌环境取2.5月龄的GFP大鼠嗅球最外层,经酶消化法分散后将细胞接种于含20%胎牛血清的DMEM/F12培养基中.定期进行光电镜观察并摄片.在培养第10天进行S-100和NGFRp75的免疫细胞化学染色鉴定,并计算OECs的纯度. 结果 培养的GFP-OECs在荧光显微镜下呈绿色强荧光,形态以双极、多突起形为主,呈S-100、NGFR p75阳性,纯度在95%以上.电镜下形态与光镜下一致,但细胞结构更为精细. 结论 本文所用方法 简便易行,可分离出高纯度的GFP-OECs,其形态学特征与生物学活性无变异.源自GFP转基因大鼠的OECs可以作为一种有效的工具细胞,广泛应用于OEC在脊髓损伤修复中的研究.

Culture and morphologic characteristics of olfactory ensheathing cells isolated from young adult GFP rat

Objective To establish methods for isolating and culturing the olfactory ensheathing cells (OECs) from young adult GIT rat to lay the foundation for repair of the spinal cord injury.Methods The OECs were dissociated from the first outer layer of the olfactory bulb of the young adult GFP rat (2.5 months old) under anatomical microscope; enzymatic digestion was performed on these cells and then they were inoculated into DMEM/F12 with 20% fetal calf serum. The morphology of OECs was regularly observed and photographied under light and electronic microscope. On the 10th culture day, the OECs were identified by the immunocytochemistry staining of S-100 and NGFRp75 and the purity (the positive rate) was calculated. Results The OECs showed strong green fluorescence under fluorescent microscope and presented morphological types of bipolar and bearing multipolar. More than 95% of the cultured cells were identified to be OECs, which expressed S-100 and NGFR p75. The cell structure revealed by electron microscope was much accordance with that by light microscope. Conclusion This method is easy to perform and high purity of GFP-OECs can be harvested with the same morphological characteristics and biological activity as general OECs. Therefore, OECs derived from the GFP transgenic rat can be effective tool cells, and widely used in studying the role of OECs in the repair of spinal cord injury