Microfluidic transfection systems

Transfection defines the process of introducing foreign biomolecules, such as DNA, RNA, liposomes, nanoparticles, antibodies and antibody fragments, into the cytoplasm or nucleus of a eukaryotic cell, with the aim of engineering or recoding cell function. This process facilitates the study of gene function, the development of personalized-targeted therapies (such as CAR-T cells) and genome editing. Conventional transfection methods are typically based on either carrier-based strategies (leveraging viral and nanoparticle-mediated systems) or the direct membrane disruption methods (such as electroporation and mechanoporation). Whilst useful, such approaches are unable to deliver large amounts of cargo in a manner that is both robust and maintains cellular viability and phenotype. Conversely, microfluidic technologies enable the precise manipulation of fluids, molecules and cells at the microscale level, suggesting potential utility in intracellular delivery. In this review, we examine new transfection technologies, highlighting the emergence of microfluidic platforms as powerful tools in overcoming the aforementioned barriers. This analysis confirms that recent advances in microfluidic electroporation, mechanoporation and sonoporation have enabled improved control over payload delivery, in turn enhancing delivery efficiencies and post-transfection cell viabilities, whilst also supporting scalable and high-throughput operation. We also discuss the roles of analytical detection and process automation in facilitating single-cell studies, high-throughput screening and therapeutic manufacturing. Finally, we highlight remaining challenges associated with clinical translation and provide a perspective on how microfluidic technologies may advance precise, efficient, and clinically relevant intracellular delivery.