Acoustofluidic sonoporation for gene delivery to human hematopoietic stem and progenitor cells
Acoustofluidic sonoporation for gene delivery to human hematopoietic stem and progenitor cells
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DOI：
10.1073/pnas.1917125117
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发表时间：
2020-05-19
影响因子：
11.1
通讯作者：
Jonas, Steven J.
中科院分区：
文献类型：
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作者：
Belling, Jason N.;Heidenreich, Liv K.;Jonas, Steven J.
基因编辑的进步正在催生新的医疗干预措施，其中患者自身的细胞被用于干细胞疗法和免疫疗法。将这些疗法转化为临床的关键限制之一是需要可扩展的技术来高效、安全地改造细胞。为了实现这一目标，诱导膜孔和渗透性的微流体策略已成为将生物分子货物输送到细胞中的有前途的技术。随着这些技术的不断成熟，需要实现临床相关细胞类型的高效、安全、无毒、快速和经济的处理。我们展示了一种声流体声孔方法，该方法基于声波对细胞膜的孔形成和透化作用，将质粒递送至永生化和原代人类细胞类型。这种声流体介导的方法可以在单通道中以 200,000 个细胞/分钟的可扩展吞吐量快速有效地将增强型绿色荧光蛋白表达质粒在细胞内递送至细胞。细胞内递送和核膜破裂的分析揭示了声流体递送和成功基因表达的机制。我们的研究表明，声流控技术是有前景的基因传递平台，也是研究膜修复的有用工具。
Advances in gene editing are leading to new medical interventions where patients' own cells are used for stem cell therapies and immunotherapies. One of the key limitations to translating these treatments to the clinic is the need for scalable technologies for engineering cells efficiently and safely. Toward this goal, microfluidic-strategies to induce membrane pores and permeability have emerged as promising techniques to deliver biomolecular cargo into cells. As these technologies continue to mature, there is a need to achieve efficient, safe, nontoxic, fast, and economical processing of clinically relevant cell types. We demonstrate an acoustofluidic sonoporation method to deliver plasmids to immortalized and primary human cell types, based on pore formation and permeabilization of cell membranes with acoustic waves. This acoustofluidic-mediated approach achieves fast and efficient intracellular delivery of an enhanced green fluorescent protein-expressing plasmid to cells at a scalable throughput of 200,000 cells/min in a single channel. Analyses of intracellular delivery and nuclear membrane rupture revealed mechanisms underlying acoustofluidic delivery and successful gene expression. Our studies show that acoustofluidic technologies are promising platforms for gene delivery and a useful tool for investigating membrane repair.