重组质粒pIRES-EGFP-BCL 11B电转染幼稚T细胞的可视化研究

Nave T Cell Transfected with Recombinant Human Vector Eukaryotic pIRES-EGFP-BCL 11B Studied by Atomic Force Microscope

将BCL11B基因插入pIRES-EGFP构建重组质粒真核表达载体pIRES-EGFP-BCL11B,采用电转染法将重组质粒转入人幼稚T细胞,转染24 h后,用原子力显微镜(AFM)观察转染前后细胞的表面形态以及生物物理性质的变化。转染72 h后,用CCK-8试剂检测幼稚T细胞的增殖情况。分别对空转的幼稚T细胞组、空载体转染组(pIRES-EGFP naked plasmid)、重组载体转染组(pIRES-EGFP-BCL 11B recombinant vector)、无电转无质粒的幼稚T细胞组进行实验。结果表明,4组幼稚T细胞的体积、高度、半宽度、粗糙度、表面颗粒大小等参数发生了变化,细胞杨氏模量以及细胞硬度也呈现很大变化,CCK-8结果显示,重组质粒pIRES-EGFP-BCL11B电转染人幼稚T细胞后影响细胞的增殖。

BCL 11B were inserted into pIRES-EGFP eukaryotic expression vector to construct recombinant pIRES-EGFP-BCL 11B plasmid. The plasmid was transfected into human naive T cell by electroporation. The atomic force microscope was used to analyze the morphology and mechanical property after transfected twenty-four hours, and CCK-8 was used to detected the proliferation of naǐve T after transfected seventy-two hours. The experiment were divided into four groups, e. g. only transfection, plRES-EGFP naked plasmid, plRES-EGFP-BCL 11B recombinant vector and no transfection or plasmid. The result showed that the morphology, ultrastructure, Young＇s modulus and stiffness of different four naǐve T cell changed greatly after transfection, and BCL 11B could promote the proliferation of naǐve T cell of human.