Cai Dong, Blair Derek, Dufort Fay J, Gumina Maria R, Huang Zhongping, Hong George, Wagner Dean, Canahan D, Kempa K, Ren Z F, Chiles Thomas C
Department of Biology, Boston College, Chestnut Hill, MA 02467, USA.
Nanotechnology. 2008 Aug 27;19(34):1-10. doi: 10.1088/0957-4484/19/34/345102.
We show herein that CNT-cell complexes are formed in the presence of a magnetic field. The complexes were analyzed by flow cytometry as a quantitative method for monitoring the physical interactions between CNTs and cells. We observed an increase in side scattering signals, where the amplitude was proportional to the amount of CNTs that are associated with cells. Even after the formation of CNT-cell complexes, cell viability was not significantly decreased. The association between CNTs and cells was strong enough to be used for manipulating the complexes and thereby conducting cell separation with magnetic force. In addition, the CNT-cell complexes were also utilized to facilitate electroporation. We observed a time constant from CNT-cell complexes but not from cells alone, indicating a high level of pore formation in cell membranes. Experimentally, we achieved the expression of enhanced green fluorescence protein by using a low electroporation voltage after the formation of CNT-cell complexes. These results suggest that higher transfection efficiency, lower electroporation voltage, and miniaturized setup dimension of electroporation may be accomplished through the CNT strategy outlined herein.
我们在此表明,在磁场存在的情况下会形成碳纳米管-细胞复合物。通过流式细胞术对这些复合物进行分析,作为监测碳纳米管与细胞之间物理相互作用的一种定量方法。我们观察到侧向散射信号增加,其幅度与与细胞相关联的碳纳米管数量成正比。即使在形成碳纳米管-细胞复合物后,细胞活力也没有显著降低。碳纳米管与细胞之间的结合足够牢固,可用于操控复合物,从而通过磁力进行细胞分离。此外,碳纳米管-细胞复合物还被用于促进电穿孔。我们观察到碳纳米管-细胞复合物存在时间常数,而单独细胞不存在,这表明细胞膜上形成了高水平的孔。通过实验,在形成碳纳米管-细胞复合物后,我们使用低电穿孔电压实现了增强型绿色荧光蛋白的表达。这些结果表明,通过本文所述的碳纳米管策略,可能实现更高的转染效率、更低的电穿孔电压以及电穿孔装置尺寸的小型化。