人衰变加速因子基因在猪血管内皮细胞中的表达

目的 通过转基因方法使猪的血管内皮细胞表达人DAF蛋白，抑制猪供人移植超急性排斥反应。方法 体外培养猪腹主动脉内皮细胞，用透射电镜方法进行鉴定。脂质体转染法将含有人DAF基因的真核表达载体PSFFV－DAV转染进猪的血管内皮细胞，经G418筛选（浓度为100mg/L)12d，获得具有抗性的克隆，进而增殖，传代，保种。通过流式细胞仪检测转染细胞DAF的表达情况。结果 转染了基因的细胞与未转染基因的细胞在流式细胞仪检测中其莹光强度分别为82.18和1.69。转染的细胞5代后其荧光强度为71.96。结论 转染了人DAF基因的细胞荧光强度明显高于对照组。导入的外源基因在细胞内能高效、稳定地表达，为转基因猪的建立奠定一定的基础。

Expression of human decay accelerating factor gene in pig aortic endothelial cells

Objective To evaluate the efficiency of inhibiting hyperacute rejections in swine to human xenotransplantation through expressing human decay accelerating factor (DAF) on the membrane of cultured pig aortic endothelial cells (PAECS) by gene transference.Methods The cultured PAECS were identified by immunoelectromicropy methods.The human DAF cDNA containing eukaryotic expression vectors PSFFV DAF were transferred into the PAECS by lipofectinmine mediated.Stable transfectants were selected using G418 (concertration 100 mg/L) and were preserved after 12 days.The expression of DAF was assayed by flow cytometry.Results The fluorescence intensity of the transfected cells and the control cells were 82.18 and 1.69 respectively in the assay by flow cytometry.The fluorescence intensity of the transfected cells could maintain 71.96 after five passages.Conclusion The fluorescence intensity of tranfected DAF expressing cells was much higher than that of control cells.Human DAF gene was stably expressed in PAECS with high efficiency.The present study provided an evidence for the further model of transgenic swine.