PAMAM-D对异种移植用外源基因转染猪精子细胞介导作用的研究  

Research on Polyamidoamine Dendrimers Mediated Foreign Gene for Xenotransplantation Transfected Pig Sperm Cells

作　　者：杨惠祥[1] 乔宝民[2] 王广有[2] 李胜芝[2] 张玥[2] 李长平[3] 徐勇[2] 马腾骧[2] 

机构地区：[1]承德医学院附属医院泌尿外科,067000 [2]天津医科大学第二医院泌尿外科天津市泌尿外科研究所 [3]天津医科大学公共卫生学院统计学教研室

出　　处：《天津医药》2010年第2期121-123,共3页Tianjin Medical Journal

基　　金：天津市应用基础研究计划项目(项目编号:07JCYBJC10700)

摘　　要：目的:探讨新型纳米材料聚酰胺-胺型树枝状聚合物PAMAM-D对异种移植用外源基因人衰变加速因子(hDAF)转染猪精子细胞的介导作用。方法:制备PAMAM-D/hDAF cDNA复合物,分为0.2、0.4、0.6、0.8和1.0μg组,每组cDNA再按照氮磷比10:1、20:1、40:1分别加入相应剂量PAMAM-D及不加PAMAM-D,对复合物进行酶切和电泳;将复合物与猪精子细胞共孵育后,原位杂交法检测hDAF对猪精子细胞的转染效率。结果:PAMAM-D能阻止PAMAM-D/DNA复合物中DNA的降解;PAMAM-D可以促进猪精子细胞对外源性DNA的摄取,其中0.4和0.6μg组的转染效率高于其他3组,0.4μg线状质粒加入PAMAM-D(氮磷比20:1)转染率为(47.5±0.2)%,是不加PAMAM-D的167%。结论:以PAMAM-D为载体可以提高外源基因对猪精子细胞的转染效率,有助于外源基因与精子细胞的稳定结合。Objective：To investigate the effect of the new nanomaterials,polyamidoamine dendrimers PAMAM-D,mediated foreign gene human decay accelerating factor （hDAF） for xenotransplantation transfected pig sperm cells.Methods：PAMAM-D/hDAF cDNA compounds were made.The compounds were divided into 0.2 μg,0.4 μg,0.6 μg,0.8 μg and 1.0 μg groups （each group adding corresponding dose of PAMAM-D in accordance with the N/P ratio 10：1,20：1,40：1）,then were digested by restriction enzymes.The compounds were incubated with washed pig sperm cells.Then the transfection efficiency was detected by in situ hybridization in the different groups.Results：The PAMAM-D molecule can prevent electrophoretic migration of DNA in the compound.After digested the compounds by restriction enzymes,DNA can not be degraded.The transfection efficiency was different in different groups.Among the total,the efficiency was higher in both groups of 0.4 μg and 0.6 μg than that of others.The top was the group of 0.4 μg linear plasmid plus PAMAM-D when the N/P ratio was 20：1（47.5% ±0.2%,167% vs control group）.Conclusion：PAMAM-D can improve the efficiency of exogenous gene transfected pig sperm cells,which can reinforce the stable binding of exogenous DNA to sperm cells.

关 键 词：精子 移植 异种 转染 基因 猪 

分 类 号：R737.25[医药卫生—肿瘤]