嗅鞘细胞的培养纯化及形态学特征

目的　建立体外培养纯化嗅鞘细胞 (Olfactoryensheathingcells,OECs)的模型 ,观察OECs生长状态和形态学变化 ,为研究神经再生获得丰富OECs来源奠定基础。方法　以原代培养的方法分离、培养成年大鼠嗅球OECs,再用阿糖胞苷抑制 ,P75抗体吸附方法纯化及Forskolin和牛垂体提取液 (bovinepituitaryextract,BPE)营养物质综合作用。根据P75免疫组化学染色结果统计所得OECs的纯度 ,同时对不同培养时期的OECs的形态进行观察。结果　 (1)此纯化方法所得的OECs纯度可达 95 %以上 ,细胞增殖速度较快 ,且细胞纯度不因培养时间的延长而明显下降。 (2 )未纯化前细胞形态多变 ,主要为巨噬细胞状、双极和多极状 ;纯化后 2～ 4天 ,OECs多呈双极或多极状 ;突起短小 ;4天后 ,OECs以突起细长的双极和三极为主 ,细胞走向一致 ,呈平行排列。结论　P75抗体吸附方法纯化效率高 ,Forskolin和BPE能够促进细胞的增殖 ,并影响细胞的形态及排布

In Vitro Culture, Purification and Morphological Properites of Olfactory Ensheathing Cells

Objective To culture and purify the olfactory ensheathing cells (OECs) in vitro and investigate the morphological changes of cultured OECs. Methods The OECs were dissociated from the olfactory bulbs of the adult rat, primarily cultured, purified with the treatment of cytosine arabinoside, and immuno-absorpted with antiserum against P 75, Forskolin and bovine pituitary extract (BPE) were added into the cultures, stimulating the OECs to divide and enrich. The purity of the OECs was evaluated according to the percentage of P 75-immunostaining cells. The morphological changes of the OECs were observed under a phase contrast microscope at different culture time. Results ① With the culturing method used in the present study, highly enriched population of OECs reached a percentage higher than 95%, and such a high purity remained when culture time prolonged. A quick proliferation of purified OECs was observed in the culture. ② The morphology of unpurified OECs varied obviously, with the majority of the cells being macr ophage-like, bipolar or multipolar. After 2～4 days of purification, bipolar and mutipolar OECs with very short processes became the main cells in the culture. When the culture time became longer than 4 days, most cells were bipolar or tripolar with long, slim processes. These cells extended unanimously and grew in parallel directions. Conclusion The methods of using P 75 antibody immunoabsorption was highly effective for the OEC purification. Forskolin and BPE are capable of promoting the proliferation of the OECs and influencing the morphology, as well as permutation of the cells.