A Flow-Through Cell Electroporation Device for Rapidly and Efficiently Transfecting Massive Amounts of Cells in vitro and ex vivo
| Autor: | Zhihong Li, Deyao Zhao, Yang Li, Qiang Cheng, Dong Huang, Zewen Wei, Xiao Zhou, Wenfeng Zhong, Yidi Wu, Zicai Liang, Mengxi Wu, Xiaoxia Wang |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Erythrocytes Green Fluorescent Proteins Microfluidics Cell Gene Expression Biology Transfection Article Cell Line Mice 03 medical and health sciences medicine Animals Viability assay Multidisciplinary Electroporation Molecular biology 030104 developmental biology medicine.anatomical_structure Cell culture Biophysics Female Low voltage Ex vivo Plasmids |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep18469 |
| Popis: | Continuous cell electroporation is an appealing non-viral approach for genetically transfecting a large number of cells. Yet the traditional macro-scale devices suffer from the unsatisfactory transfection efficiency and/or cell viability due to their high voltage, while the emerging microfluidic electroporation devices is still limited by their low cell processing speed. Here we present a flow-through cell electroporation device integrating large-sized flow tube and small-spaced distributed needle electrode array. Relatively large flow tube enables high flow rate, simple flow characterization and low shear force, while well-organized needle array electrodes produce an even-distributed electric field with low voltage. Thus the difficulties for seeking the fine balance between high flow rate and low electroporation voltage were steered clear. Efficient in vitro electrotransfection of plasmid DNA was demonstrated in several hard-to-transfect cell lines. Furthermore, we also explored ex vivo electroporated mouse erythrocyte as the carrier of RNA. The strong ability of RNA loading and short exposure time of freshly isolated cells jointly ensured a high yield of valid carrier erythrocytes, which further successfully delivered RNA into targeted tissue. Both in vitro and ex vivo electrotransfection could be accomplished at high cell processing speed (20 million cells per minute) which remarkably outperforms previous devices. |
| Databáze: | OpenAIRE |
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