Microfluidic platforms for the transient transfection of mammalian cells: recent developments and challenges

Transient transfection is a widely used technique for the temporary introduction of nucleic acids into cells, enabling short-term gene expression without permanent genomic integration. Compared to stable transfection, it is faster, more cost-effective, and flexible, though expression remains time-limited. Interest in this method has grown substantially in fields requiring small sample volumes or high parallelization, including personalized medicine, gene therapy, and high-throughput screening. Microfluidic systems offer strong potential due to their superior automation and reproducibility. Methods such as electroporation, mechanoporation, and nanoparticle-based delivery have increasingly been translated into microfluidic platforms and are undergoing continuous optimization. This review summarizes current approaches, focusing on mRNA and plasmid DNA delivery, and discusses key challenges for integration into industrial biopharmaceutical production and gene therapy workflows.