【摘要】 为寻求使猪主动脉血管内皮细胞增殖寿命延长的方法,本试验将含有人类端粒酶催化亚基端粒酶逆转录酶(Human telomerase reverse transcriptase,hTERT)基因的真核表达质粒pCI-neo-hTERT通过脂质体介导法导入猪主动脉血管内皮细胞(Porcine arterial endothelial cell,PAEC)。对转染后的血管内皮细胞进行Ⅷ因子和CD34鉴定,利用PCR-ELISA方法检测hTERT导入后细胞端粒酶活性的表达情况,探讨了转染后细胞的生长特性。结果显示,转染后的细胞Ⅷ因子和CD34均呈阳性,证明转染后细胞仍表达血管内皮细胞的特异性蛋白;导入hTERT后,细胞有较高的端粒酶活性,增殖寿命较对照细胞延长了20代。表明hTERT导入细胞能重建细胞端粒酶活性,从而延长细胞在体外培养的寿命。更多还原

【Abstract】 To elongate the proliferation life-span of porcine arterial endothelial cell(PAEC) in vitro.The eukaryotic expression plasmid pCI-neo-hTERT was transfected into PAEC by lipofection.FactorⅧ related antigen and CD34 of PAEC were identified after transfection by immunohistochemical method;we also detected telomerase activity of PAEC after transfection by PCR-ELISA.The results showed that Factor Ⅷ related antigen and CD34 in PAEC was positive after transfection.The PAEC after transfection expressed the vascular endothelial cell-specific protein still;Telomerase activity in PAEC was higher by hTERT introduction,the cell proliferation life-span was elongated to 20 population doublings compared to control.Our research indicated that telomerase activity can be activated in PAEC,thus the cell proliferation life-span was elongated in vitro.更多还原