文章摘要
荣宁,汪曣,范真真.基质刚性影响声孔效应介导的单细胞基因转染[J].,2020,39(5):764-768
基质刚性影响声孔效应介导的单细胞基因转染
Matrix rigidity affects sonoporation mediated gene delivery on single cell
投稿时间:2019-11-27  修订日期:2020-09-02
中文摘要:
      实验研究了基质刚性对单细胞质粒DNA转染效果的影响。实验采用高声压短脉冲(0.45MPa,10μs)条件的超声对培养在不同硬度凝胶基质(软的凝胶基质:0.2kPa,硬的凝胶基质:40kPa)上的力学敏感细胞NIH 3T3进行质粒DNA转染实验。实验结果表明,培养在硬的凝胶基质上的细胞,质粒DNA转染效率明显高于培养在软的凝胶基质上的细胞。进一步对质粒DNA进行荧光示踪可知培养在不同刚性基质上的细胞导入质粒DNA的方式不同。当细胞被培养在硬的凝胶基质上时,通过声致穿孔产生的小孔进入细胞内的质粒DNA更多,而培养在软的凝胶基质上的细胞,更多的质粒DNA可以通过非声致穿孔作用,例如内吞方式导入细胞。 细胞骨架蛋白分布规律表明,硬的凝胶基质上培养的细胞内有更多的F肌动蛋白微丝,可以更好地支撑起细胞的铺展形态,相对不容易发生内吞作用。而软的凝胶基质上培养的细胞内F肌动蛋白则更多以球形状态存在,细胞形貌骗向圆形,此时更容易发生胞吞作用。
英文摘要:
      This research study the effects of matrix rigidity on single-cell plasmid DNA transfection. NIH 3T3 cells were cultured on hydrogels with different rigidities (soft gel substrate: 0.2 kPa, hard gel substrate: 40 kPa), and a high-pressure short-pulse (0.45 MPa, 10 μs) ultrasound was used for gene transfection experiment. The experimental results showes that the plasmid DNA transfection efficiency of cells cultured on a hard substrate is ??significantly higher than that of cells cultured on a soft gel matrix. Further fluorescence tracing of plasmid DNA showed that DNA can be intracellular uptake through separate ways for cells cultured on substrates with different rigidities. When cells are cultured on a rigid substrate, more plasmids enter the cytoplasm through cell membrane pores generated by sonoporation, while for cells cultured on a soft substrate, more plasmid DNA can be intracellular uptake through other ways, such as endocytosis. More F-actin microfilaments in cells cultured on rigid substrate supported the spreading morphology, while F-actin turned out to be clusters in cells on soft substrate which made it much easier for endocytosis.
DOI:10.11684/j.issn.1000-310X.2020.05.015
中文关键词: 基质刚性  超声  微泡  基因转染
英文关键词: matrix rigidity  ultrasound  microbubble  gene transfection
基金项目:(11874280),中国科学院声学研究所声场与声信息国家重点实验室重点项目(SKLA201910),天津市自然科学基金项目(19JCZDJC33600)
作者单位E-mail
荣宁 天津大学 生物医学工程 天津 rongning@tju.edu.cn 
汪曣 天津大学 生物医学工程 天津 wangyan@tju.edu.cn 
范真真 天津大学 生物医学工程 天津 zhenzhen.fan@tju.edu.cn 
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