论文部分内容阅读
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。
Objective To investigate the optimization of culture and purification of human embryonic olfactory ensheathing cells (OECs). Methods The aborted fetus was removed from 3 to 5 months and the olfactory bulb was removed without 6 bacteria. The soft membrane tissues and blood vessels on the surface of the olfactory bulb were dissected carefully and dissected. After digestion, conditioned medium was used to prepare 10-cell / ml cell suspension , Purification and purification of serum-free purified liquid using differential adherence method combined with cytarabine inhibition method, and observation of different culture periods 75 and immunization with PNGFR (Low Affinity Neurotrophic Factor Receptor) and FBN (Fibronectin) Fluorescent staining identification. Results The preparation method could effectively remove fibroblasts and obtain 90% pure olfactory ensheathing cells. Olfactory ensheathing cells were oval-shaped somatic cells and slender synapses under microscope. Most of the cells were bipolar or polypolar, Immunofluorescence staining NGFRP positive cells percentage of 80% -90%. Conclusion The culture purification method is feasible and can be obtained in line with the requirements of clinical cell transplantation for cell preparation.