人胚胎嗅球嗅鞘细胞的制备  

The preparation of human embryo olfactory bulb olfactory ensheathing cells

作　　者：肖娟[1] 王洪美[1,2] 陈琳[1,2] 任玉水[1] 姜晓荣[1] 黄红云[1,2] 李莹[1] 赵艳秀[1] 

机构地区：[1]北京市虹天济神经科学研究院 [2]北京康复中心神经外科,北京100144

出　　处：《解剖科学进展》2011年第3期276-279,共4页Progress of Anatomical Sciences

摘　　要：目的探讨人胚胎嗅鞘细胞(OECs)培养与纯化的优化方法。方法取3～5个月流产胚胎,无6菌条件下取出嗅球,解剖镜下仔细剥离嗅球表面的软膜组织和血管,消化后用条件培养基制成密度为10个细胞/ml细胞悬液,利用差速贴壁法结合阿糖胞苷抑制法进行无血清纯化液细胞纯化培养,对不同培养时期进行观察75和行PNGFR(低亲和力神经营养因子受体)和FBN(纤维粘连蛋白)免疫荧光染色鉴定。结果此制备法可以有效去除成纤维细胞,获得纯度90%嗅鞘细胞,显微镜下观察嗅鞘细胞有卵圆形的胞体和细长的突触,细胞多呈75为双极或多极形,免疫荧光染色NGFRP阳性细胞百分数为80%-90%。结论本培养纯化方法可行,可获得符合临床细胞移植治疗要求细胞制剂。Objectives To inquire into the optimized method in culturing and puring human embryo olfactory bulb olfactory ensheathing cells. Methods The olfactory bulbs of 3 to 5 months＇ human embryo were taken out and aborted into sterilized condition, the meninx vasculosa tissue and blood vessels were peeled off from the surface of the bulbs under anatomical lens carefully. The bulbs were preparated into cell suspension using conditioned medium with density of 106 cells per milliliter after digesting. The cells were observed in different culture stage and immunofluorescence staining was performed with anti-P^75NGFR （low-affinity neurotrophic factor receptor） and anti-FBN （fibronectin） for identification. Results This method of preparation can remove fibroblasts effectively and obtain 90% of olfactory ensheathing cells. The olfactory ensheathing cells were observed with ovalis cell bodies and bistoury synapses and bipolar or muhipolar branches under microscope, and showed 80%-90% of P^75NGFR positive and FBN positive by immunofluorescence staining. Conclusions This culture purification method is available in obtaining cells for clinical cells transplantation therapy need.

关 键 词：人嗅鞘细胞 纯化 培养 低亲和力神经营养因子受体 纤维粘连蛋白 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]