【摘要】 目的评价以纳米粒子ORMA40为载体介导生存素基因转染嗅神经鞘细胞的效率.方法合成ORMA40;扩增、分离并纯化人类生存素基因的质粒pS和mS;制备ORMA40-pS/mS复合体pSO和mSO;分离、培养、纯化并鉴定嗅神经鞘细胞(olfactory nerve ensheathing cells,ONECs);分别采用pS、pSO、mS或mSO转染ONECs.结果电镜表明,水相ORMA40分散好、平均直径为20 nm;电泳表明,mSOC和pSOC不被DNase 1消化;免疫细胞染色表明,mS、pSO和mSO可转染50%以上的ONECs;统计分析表明,pSO和mSO转染ONECs的效率比pS转染者高(P<0.01)、与mS转染者相当(P>0.05).结论以纳米粒子ORMA40为载体可高效地介导生存素基因转染ONECs.更多还原

【Abstract】 Objective To explore the effectiveness of nanoparticle ORMA40 as a vector for delivery of survivin gene to olfactory nerve ensheathing cells(ONECs).Methods ORMA40 nanoparticles(ORMA40) was synthezed,and plasmid survivin(pS and mS)waspropagated,isolated and purifyed to form ORMA40-survivin complexes(pSO and mSO);ONECs was isolated,cultured,purifyed and identifyed to transfect ONECs with pS,mS,pSO or mSO.Results Transmission electron microscopy showed that ORMA40 were highly monodispersed in the aqueous solution with the average size of 20 nm;agarose gel electrophoresis showed that both mSO and pSO were free from digestion of DNase 1;immunostaining showed that mS,pSO and mSO can transfect more than 50 % of ONECs;and statistical analysis showed that the transfection effectiveness of pSO and mSO was higher than that of pS(P<0.01)and as the same as that of mS(P >0.05).Conclusion Nanoparticle ORMA40,as a vector,can effectively mediate the delivery of survivin gene to ONECs.更多还原