阳离子脂质体介导的外源基因转染小鼠脾细胞的方法学探讨

Methodological Investigation of Foreign Gene Transferring Mouse Spleen Cells Mediated by Cationic Liposomes

【目的】探讨高效阳离子脂质体介导外源基因转染体外生长的小鼠脾细胞的方法。【方法】用新型阳离子脂质体DOSPER包裹携带lacZ报告基因的质粒 pCAGGS lacZ按 3种方法转染小鼠脾细胞 ,①常规方法直接转染 ;②脾细胞经刀豆球蛋白A(ConA)活化后再按常规方法转染 ;③用黏附辅助脂质体法 (adhesion assistedlipofection ,AAL)转染脾细胞。转染效率用X gal染色法测定。【结果】上述 3种方法的转染效率依次为 <0 0 10 ,0 0 5 7及 0 199,经方差分析 ,3种方法转染效率的差异有显著性 (P <0 0 1,n =3)。【结论】AAL法介导外源基因转染体外生长的小鼠脾细胞的转染效率最高。

Objective To investigate the high efficient methods of foreign gene transferring mouse spleen cells cultured in vitro mediated by cationic liposomes. Methods Mouse spleen cells were transfected with Lac Z reporter gene harbored plasmid pCAGGS-lacZ coated by cationic liposome DOSPER using the following three alternative methods: First, the spleen cells were transfected directly with plasmids/liposome complexes using conventional procedure; Second, Concanavalin A (Con A) stimulated spleen cells were transfected using routine method; Third, spleen cells were transfected using adhesion-assisted lipofection (AAL) method. The transfection efficiency was determined by X-gal staining method. Results The transfection efficiency of the three methods were <0.010,0.057 and 0.199 respectively. Statistical results of SNK Test showed significant differences (P<0.01, n=3) among the three groups. Conclusion AAL mediated foreign gene transferring the in vitro cultured mouse spleen cells shows the highest efficiency.