In vitro and ex vivo intracellular delivery methods hold the key for releasing the full potential of tissue engineering, drug development, and many other applications. In recent years, there has been significant progress in the design and implementation of intracellular delivery systems capable of delivery at the same scale as viral transfection and bulk electroporation but offering fewer adverse outcomes. This review strives to examine a variety of methods for in vitro and ex vivo intracellular delivery such as flow‐through microfluidics, engineered substrates, and automated probe‐based systems from the perspective of throughput and control. Special attention is paid to a particularly promising method of electroporation using micro/nanochannel based porous substrates, which expose small patches of cell membrane to permeabilizing electric field. Porous substrate electroporation parameters discussed include system design, cells and cargos used, transfection efficiency and cell viability, and the electric field and its effects on molecular transport. The review concludes with discussion of potential new innovations which can arise from specific aspects of porous substrate‐based electroporation platforms and high throughput, high control methods in general.

中文翻译：

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用于体外细胞内递送的高通量和高度可控的方法

体外和离体细胞内递送方法是释放组织工程、药物开发和许多其他应用的全部潜力的关键。近年来，细胞内递送系统的设计和实施取得了重大进展，能够以与病毒转染和批量电穿孔相同的规模进行递送，但不良后果更少。本综述致力于从通量和控制的角度研究体外和离体细胞内递送的各种方法，例如流通微流体、工程基质和基于自动化探针的系统。特别关注使用基于微/纳米通道的多孔基质的一种特别有前景的电穿孔方法，该方法将小片细胞膜暴露于透化电场。讨论的多孔基质电穿孔参数包括系统设计、使用的细胞和货物、转染效率和细胞活力、电场及其对分子运输的影响。该综述最后讨论了基于多孔基质的电穿孔平台的特定方面和高通量、高控制方法的潜在新创新。