Kim Min Gon, Yoon Sangpil, Chiu Chi Tat, Shung K Kirk
Department of Biomedical Engineering, University of Southern California, Los Angeles, California, USA.
Department of Biomedical Engineering, University of Southern California, Los Angeles, California, USA.
Ultrasound Med Biol. 2018 Mar;44(3):622-634. doi: 10.1016/j.ultrasmedbio.2017.11.005. Epub 2017 Dec 25.
Manipulation of cellular functions and structures by introduction of genetic materials inside cells has been one of the most prominent research areas in biomedicine. High-frequency ultrasound acoustic-transfection has recently been developed and confirmed by intracellular delivery of small molecules into HeLa cells at the single-cell level with high cell viability. After we proved the concept underlying the acoustic-transfection technique, treatment conditions for different human cancer cell lines have been intensively investigated to further develop acoustic-transfection as a versatile and adaptable transfection method by satisfying the requirements of high-delivery efficiency and cell membrane permeability with minimal membrane disruption. To determine optimal treatment conditions for different cell lines, we developed a quantitative intracellular delivery score based on delivery efficiency, cell membrane permeability and cell viability after 4 and 20 h of treatment. The intracellular delivery of macromolecules and the simultaneous intracellular delivery of two molecules under optimal treatment conditions were successfully achieved. We found that DNA plasmid was delivered by acoustic-transfection technique into epiblast stem cells, which expressed transient mCherry fluorescence.
通过将遗传物质引入细胞内部来操纵细胞功能和结构，一直是生物医学领域最突出的研究领域之一。高频超声声转染技术最近得到了发展，并通过在单细胞水平将小分子高效递送至HeLa细胞内且保持高细胞活力得到了证实。在我们证明了声转染技术的基本原理之后，为了进一步将声转染发展成为一种通用且适应性强的转染方法，通过满足高递送效率和细胞膜通透性要求且使细胞膜损伤最小化，我们对不同人类癌细胞系的处理条件进行了深入研究。为了确定不同细胞系的最佳处理条件，我们基于处理4小时和20小时后的递送效率、细胞膜通透性和细胞活力，制定了一个定量细胞内递送评分标准。在最佳处理条件下，成功实现了大分子的细胞内递送以及两种分子的同时细胞内递送。我们发现，通过声转染技术可将DNA质粒递送至表达瞬时mCherry荧光的上胚层干细胞中。
