新生大鼠嗅球嗅鞘细胞的纯化方法研究

目的 探讨新生大鼠嗅球嗅鞘细胞（OECs）的体外培养和纯化方法，为研究脊髓损伤后神经再生获得丰富OECs来源奠定基础。方法 用原代培养的方法分离、培养新生大鼠嗅球OECs，比较p75抗体吸附、阿糖胞苷抑制、差速贴壁三种方法的纯化效果，根据GFAP免疫细胞化学染色结果统计所得OECs的纯度，同时在相差显微镜下对不同培养时期的OECs的形态进行观察。结果 体外培养的新生大鼠嗅球OECs主要为双极或三级细胞，其突起细长。未经纯化的OECs则成纤维细胞生长迅速而占优势，三种纯化方法均能使接种14d后的OECs纯度均值大于75％。p75抗体吸附法和阿糖胞苷抑制法的纯化率均值略高于差速贴壁法。结论 新生大鼠嗅球OECS的原代培养中细胞纯化是必要的；在脊髓损伤再生研究中，较之p75抗体吸附法和阿糖胞苷抑制法，差速贴壁法是一种简单、经济、实用的OECs纯化方法。

Purification of Olfactory Ensheathing Cells from Neonatal Rat Olfactory Bulbs

Objective To culture and purify the olfactory ensheathing cells(OECs) in vitro,and to investigate the morphological features and the immunocytochemical properties of the cultured OECs.Methods The OECs were dissociated from the neonatal rat olfactory bulbs.The primarily cultured OECS were purified with the treatment of arabinoside cytosine(Ara-C),or immune-absorption with anti-p75 antibodies,or a method based on the differing rates of attachment of the various harvestedcell types.The purity of the OECs was evaluated according to the percentage of GFAP-immunostaining cells.The morphological changes of the OECs were observed under a phase contrast microscope at different culture time.Results The OECs display a very characteristic morphological appearance.Most of OECs are bipolar or tripolar with long and thin processes.In the unpurified group the rapidly proliferating fibroblasts were in the majority after 7 days in culture.Whereas in other three puritied groupes the purity of OECs was estimated to be over 75% after 14 days in culture.The OECs purification by the method based on the fiffering rates of attachment of the various harvested cell types was a litter inefficient than those methods of using anti-p75 antibody or Ara-C.Conclusion The puritification is necessary in Primary culture of olfactory ensheathing cells from neonatal rat olfactory bulbs.The method of purification for OECs based on the differing rates of attachment of the various cell types is simple.inexpensive and practical in the research of the regeneration after spinal cord injury.