Salari Alinaghi, Appak-Baskoy Sila, Coe Imogen R, Abousawan John, Antonescu Costin N, Tsai Scott S H, Kolios Michael C
Institute for Biomedical Engineering, Science and Technology (iBEST), Toronto, ON M5B 1T8, Canada and Biomedical Engineering Graduate Program, Ryerson University, Toronto, ON M5B 2K3, Canada.
Institute for Biomedical Engineering, Science and Technology (iBEST), Toronto, ON M5B 1T8, Canada and Department of Chemistry and Biology, Ryerson University, Toronto, ON M5B 2K3, Canada.
Lab Chip. 2021 May 4;21(9):1788-1797. doi: 10.1039/d0lc01303j.
Biological research and many cell-based therapies rely on the successful delivery of cargo materials into cells. Intracellular delivery in an in vitro setting refers to a variety of physical and biochemical techniques developed for conducting rapid and efficient transport of materials across the plasma membrane. Generally, the techniques that are time-efficient (e.g., electroporation) suffer from heterogeneity and low cellular viability, and those that are precise (e.g., microinjection) suffer from low-throughput and are labor-intensive. Here, we present a novel in vitro microfluidic strategy for intracellular delivery, which is based on the acoustic excitation of adherent cells. Strong mechanical oscillations, mediated by Lamb waves, inside a microfluidic channel facilitate the cellular uptake of different size (e.g., 3-500 kDa, plasmid encoding EGFP) cargo materials through endocytic pathways. We demonstrate successful delivery of 500 kDa dextran to various adherent cell lines with unprecedented efficiency in the range of 65-85% above control. We also show that actuation voltage and treatment duration can be tuned to control the dosage of delivered substances. High viability (≥91%), versatility across different cargo materials and various adherent cell lines, scalability to hundreds of thousands of cells per treatment, portability, and ease-of-operation are among the unique features of this acoustofluidic strategy. Potential applications include targeting through endocytosis-dependant pathways in cellular disorders, such as lysosomal storage diseases, which other physical methods are unable to address. This novel acoustofluidic method achieves rapid, uniform, and scalable delivery of material into cells, and may find utility in lab-on-a-chip applications.
生物学研究和许多基于细胞的疗法都依赖于将货物材料成功递送至细胞内。体外环境下的细胞内递送是指为实现材料快速高效地跨质膜运输而开发的多种物理和生化技术。一般来说，省时的技术（如电穿孔）存在异质性且细胞活力低的问题，而精确的技术（如显微注射）则存在通量低且劳动强度大的问题。在此，我们提出一种用于细胞内递送的新型体外微流控策略，该策略基于对贴壁细胞的声激发。微流控通道内由兰姆波介导的强烈机械振荡促进了不同大小（如3 - 500 kDa，编码增强型绿色荧光蛋白的质粒）的货物材料通过内吞途径被细胞摄取。我们证明了500 kDa葡聚糖以高达65 - 85%的前所未有的效率成功递送至各种贴壁细胞系，高于对照组。我们还表明，可以调节驱动电压和处理持续时间来控制递送物质的剂量。高活力（≥91%）、对不同货物材料和各种贴壁细胞系的通用性、每次处理可扩展至数十万细胞、便携性以及操作简便性是这种声流体策略的独特特征。潜在应用包括通过内吞依赖途径靶向细胞疾病，如溶酶体贮积病，这是其他物理方法无法解决的。这种新型声流体方法实现了将材料快速、均匀且可扩展地递送至细胞内，可能在芯片实验室应用中发挥作用。
