Presentation Information
[10a-C309-4]Nanotube Stamping for Therapeutic protein and Mitochondria Delivery into Suspension Cells
〇(M2)Jizhou Jin1, Bingfu Liu1, Kazuhiro Oyama1, Takeo Miyake1 (1.Waseda Univ.)
Keywords:
Bioelectronics,Nanotube Stamping,Intracellular Delivery
In regenerative medicine, cell therapy often relies on suspension cells, creating a critical need for efficient methods to deliver multiple cargoes into these cells. As a first step towards this goal, our previous nanotube (NT) stamping platform enabled efficient delivery into adherent cells [1-3], but delivery into the critically important suspension cells—a major target in therapeutic applications—has not yet been realized. Here, we present a versatile NT stamping platform adapted for high-efficiency, size-independent delivery into suspension cells, such as THP-1, without compromising viability. Our platform consists of a vertically aligned gold nanotube array integrated onto a glass stamp, which interfaces with cells under microscopic control. By optimizing NT geometry and insertion depth, we achieved efficient intracellular injection of fluorescent dyes, proteins (GFP, green fluorescent protein; LOx, lactate oxidase, Cas9/sgRNA), and mitochondria into THP-1 cells immobilized using a gentle surface-coating method, with delivery efficiencies exceeding 90% and cell viability above 85%. Unlike conventional methods such as electroporation, our approach maintains cell functionality and enables precise, reagent-free delivery. Notably, the platform’s ability to reversibly extract and inject intracellular fluid enabled the direct transfer of stem cell–derived mitochondria into suspension cells, enhancing ATP production per cell by twofold for at least one week. These results highlight the potential of our system to overcome the inherent limitations of conventional delivery methods and to accommodate a wide range of biomolecular cargo.