Abstract：
Efficient and convenient delivery of exogenous molecules into cells is important for cell biology research. However, many intracellular delivery methods require carrier-mediated or physical field assistance, complicating the delivery process. Here, a general, simple, and effective method for in situ single-cell intracellular delivery is reported. A solution containing digitonin and cargo is precisely applied to single cells using a microfluidic probe. Digitonin binds to cholesterol in the plasma membrane to induce perforation, and the cargo enters the cell through the pore. By optimizing parameters, propidium iodide (0.67 kDa) and FITC-dextran (10, 40, and 150 kDa) can be successfully introduced into single cells within 3 min while maintaining cell viability. To prove the potential of this method for cell research, we delivered cytochrome C (13 kDa) and cyclophilin A (18 kDa) into cells by this method. The delivered cytochrome C successfully induces cell apoptosis by activating the caspase pathway, and cyclophilin A performs an antioxidant effect in the cells, which may enhance the drug resistance of glioma cells. It is believed that this method will be an attractive tool for single-cell intracellular delivery.
摘要：
高效、便捷地将外源分子递送到细胞中对于细胞生物学研究具有重要意义。然而,许多细胞内递送方法需要载体介导或物理场辅助，使交付过程复杂化。这里,一个将军,简单,报道了原位单细胞细胞内递送的有效方法。使用微流体探针将含有毛地黄皂苷和货物的溶液精确地施加到单细胞。洋地黄皂苷与质膜中的胆固醇结合以诱导穿孔，货物通过孔进入细胞。通过优化参数，碘化丙啶（0.67kDa）和FITC-葡聚糖（10、40和150kDa）可以在3分钟内成功引入单个细胞，同时保持细胞活力。为了证明这种方法在细胞研究中的潜力，我们通过这种方法将细胞色素C（13kDa）和亲环素A（18kDa）递送到细胞中。传递的细胞色素C通过激活caspase途径成功诱导细胞凋亡，亲环蛋白A在细胞中发挥抗氧化作用，这可能会增强神经胶质瘤细胞的耐药性。相信该方法将是用于单细胞细胞内递送的有吸引力的工具。
