摘要
Intracellular delivery of therapeutic biomolecules represents a fundamental prerequisite for cell-based therapies and precision medicine, yet existing delivery methods present critical limitations. Viral vectors, while effective, pose safety risks including immunogenicity and insertional mutagenesis. Bulk electroporation offers a non-viral alternative but suffers from high cytotoxicity, heterogeneous electric field distributions, and poor efficiency in primary cells due to excessive voltage requirements and uncontrolled Joule heating.
Microfluidic electroporation exploits microscale physics to decouple transfection efficiency from cell viability. By reducing electrode spacing to micrometers, these platforms achieve necessary field strengths at voltages below 50 V, minimizing Joule heating and electrolysis byproducts that plague bulk methods. Static platforms, including nanostructure-assisted designs, provide subcellular precision through localized field enhancement and real-time impedance monitoring, enabling mechanistic investigation of pore formation dynamics. Continuous-flow systems transform electroporation into a scalable manufacturing process, achieving throughputs of 108–109 cells per minute required for clinical cell therapy production while maintaining viabilities above 90% through hydrodynamic focusing and optimized channel geometries.
Despite these engineering advances, systematic benchmarking against Current Good Manufacturing Practice (cGMP)-compliant commercial electroporators reveals critical translational barriers: reliance on research-grade polydimethylsiloxane instead of medical-grade thermoplastics, open manual workflows incompatible with sterile closed-system requirements, lack of validated process control protocols, and insufficient biological verification beyond transient fluorescent protein expression. Furthermore, cargo-specific constraints, including nuclear transport requirements for plasmid DNA versus ribonucleoprotein complexes and distinctions between transient mRNA expression versus permanent CRISPR/Cas9 genomic integration, demand fundamentally different optimization strategies rarely addressed in device-focused studies. Establishing microfluidic electroporation as a viable clinical platform requires integrated manufacturing modules coupling electroporation with upstream buffer exchange and downstream cell sorting, along with implementation of real-time process analytical technology and closed-loop artificial intelligence-driven control. Early regulatory engagement to establish Drug Master File pathways will enable broad therapeutic applications.
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