Flexible Mechanoporation Chips for High-Throughput Intracellular Delivery Based on Controlled Pneumatic Microvalve Array.

Mechanoporation-based intracellular delivery has emerged as an effective technique for transporting materials into living cells through the application of mechanical loading and deformation. Although this technique holds significant potential for large-scale cell manufacturing without the need for additional carriers, there exist several challenges, including a high degree of size dependence, inconsistencies due to cellular heterogeneity, and the risk of channel clogging and cell damage. Here, we developed a flexible mechanoporation chip system that integrated a three-layer pneumatic microvalve array for high-throughput intracellular delivery. Both our simulation data and experimental results indicated that it could minimize cell damage and enhance delivery efficiency through volume exchange and molecular convection. The adaptive deformation design of the microvalve array allowed it to accommodate variations in the geometric sizes and mechanical properties of cell populations, thereby optimizing intracellular delivery. Furthermore, we demonstrated that the flexible mechanoporation chip system could effectively deliver various biomolecules, including drugs, mRNA, and plasmid DNA, into diverse cell types like mouse embryonic fibroblasts (MEFs), adipose-derived stem cells (ASCs), and primary T cells. This flexible mechanoporation chip platform presents a promising tool for efficient, high-throughput, and low-damage mechanical transfection in biomanufacturing, cell therapy and regenerative medicine.