Intracellular Photothermal Delivery for Suspension Cells Using Sharp Nanoscale Tips in Microwells
| Autor: | Tingyi Leo Liu, Pei-Shan Chung, Daniel L. Clemens, Jason S. Hong, Michael A. Teitell, Tianxing Man, Pei-Yu Chiou, Bai-Yu Lee, Chuanzhen Zhao, Cong Wu, Yu Ting Chow, Alexander N. Patananan, Paul S. Weiss, Xiongfeng Zhu, Ximiao Wen, Emma R. Dawson |
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| Rok vydání: | 2019 |
| Předmět: |
Cell Survival
medicine.medical_treatment Finite Element Analysis Green Fluorescent Proteins Intracellular Space General Physics and Astronomy Nanotechnology 02 engineering and technology 010402 general chemistry 01 natural sciences beta-Lactamases Suspensions Cancer immunotherapy Cell Line Tumor medicine Humans General Materials Science Suspension (vehicle) Nanoscopic scale chemistry.chemical_classification Chemistry Lasers Biomolecule General Engineering Hyperthermia Induced Phototherapy Photothermal therapy 021001 nanoscience & nanotechnology 0104 chemical sciences Nanoparticles 0210 nano-technology Intracellular Gravitation |
| Zdroj: | ACS Nano. 13:10835-10844 |
| ISSN: | 1936-086X 1936-0851 |
| DOI: | 10.1021/acsnano.9b06025 |
| Popis: | Efficient intracellular delivery of biomolecules into cells that grow in suspension is of great interest for biomedical research, such as for applications in cancer immunotherapy. Although tremendous effort has been expended, it remains challenging for existing transfer platforms to deliver materials efficiently into suspension cells. Here, we demonstrate a high-efficiency photothermal delivery approach for suspension cells using sharp nanoscale metal-coated tips positioned at the edge of microwells, which provide controllable membrane disruption for each cell in an array. Self-aligned microfabrication generates a uniform microwell array with three-dimensional nanoscale metallic sharp tip structures. Suspension cells self-position by gravity within each microwell in direct contact with eight sharp tips, where laser-induced cavitation bubbles generate transient pores in the cell membrane to facilitate intracellular delivery of extracellular cargo. A range of cargo sizes were tested on this platform using Ramos suspension B cells with an efficiency of84% for Calcein green (0.6 kDa) and45% for FITC-dextran (2000 kDa), with retained viability of96% and a throughput of100 000 cells delivered per minute. The bacterial enzyme β-lactamase (29 kDa) was delivered into Ramos B cells and retained its biological activity, whereas a green fluorescence protein expression plasmid was delivered into Ramos B cells with a transfection efficiency of58%, and a viability of89% achieved. |
| Databáze: | OpenAIRE |
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