成年鼠嗅鞘细胞的纯化培养与免疫组化观察

Purification of Adult Rat Olfactory Ensheathing Cells and Observation of Cell's Morphology and Immunohistochemistry

目的 :对文献报道的嗅鞘细胞 (OECs)纯化方法进行改进 ,为基因治疗和细胞移植修复脊髓损伤实验研究提供高纯度的种子细胞。方法 :本实验综合使用了阿糖胞苷、BPE及NGFR抗体纯化了原代培养的成年Wistar大鼠的OECs,并对纯化的OECs细胞免疫特性进行观察。结果 :本实验得到OECs纯度可达 98% ,其特性与文献报道一致。结论 :本实验综合使用了阿糖胞苷、BPE及NGFR抗体纯化了原代培养的成年Wistar大鼠OECs,纯度可达 98%。而且培养周期大大缩短 ,节约了时间 ,为进一步的实验研究奠定了基础。

Objective:To improve the method of culture and purification of olfactory ensheathing cells (OECs) for gene therapy or cell transplantation in injuried spinal cord repair.Method:Olfactory nerve and glomerular layers from neonatal rats bulbles were dissociated and cultured in DMEM/F-12 medium,and OECs were incubated in the medium including cytosine arabinoside for 24-48h and BPE for 5 days. When the cells were confluent, they were trypsinized and purified by immunoaffinity of anti-NGFR and the cells immunity properties were observed.Result:Two weeks later the OECs dominated the cultures and revealed two types of form:the fusiform and multipolar cells,they were stained with S100 protein and NGFR positively. Conclusion:The OECs can be purified to 98 percent purity by the method and the period of purification is shortened.