An Advanced Microfluidic Electroporation System for Efficient Gene Delivery in Patient-Derived Cells

Patient-derived cells are crucial in personalized medicine for precise disease modeling, efficient drug screening, and tailored therapeutic development. Traditional viral methods for gene insertion pose risks, such as viral genomic integration and mutations. Nonviral methods like electroporation also face challenges, including cytotoxicity from proprietary buffers, high voltage requirements, and inconsistent transfection efficiencies due to cell size disparities. Additionally, affinity-based cell pre-purification complicates clinical translation.
We developed an integrated electroporation system that utilizes an ultra-high throughput vortex cell purification method. This platform efficiently purifies, permeabilizes, and delivers genetic constructs to primary cells using a microfluidic chip and microscale electrode system. The chip employs size-based vortex purification for uniform cell trapping across 144 parallel trapping chambers. Interdigitated microelectrodes generate localized electric fields of 1.5 kV/cm with low input voltages (<40 V), enhancing throughput by 3.6-fold over previous prototypes while maintaining comparable performance.
The system's rapid solution exchange allows for in situ customization of buffer compositions, improving transfection outcomes compared to conventional buffers. We demonstrated enhanced gene delivery efficiency in human mammary fibroblast primary cells using an optimized electroporation buffer, achieving up to 8-fold higher efficiency than DPBS and matching lipofection performance (>80%). Additionally, we achieved robust protein production from synthetic mRNA in HMF transfections, demonstrating the system's potential.
This novel electroporation system is being validated with circulating tumor cells from breast cancer patients to demonstrate its potential for personalized medicine.