Chang Lingqian, Howdyshell Marci, Liao Wei-Ching, Chiang Chi-Ling, Gallego-Perez Daniel, Yang Zhaogang, Lu Wu, Byrd John C, Muthusamy Natarajan, Lee L James, Sooryakumar Ratnasingham
NSEC Center for Affordable Nanoengineering of Polymeric Biomedical Devices, Ohio State University, Columbus, OH 43209, USA.
Comprehensive Cancer Center, The Ohio State University, Columbus, OH, 43209, USA. Division of Hematology, The Comprehensive Cancer Center, Ohio State University, Columbus, OH, 43209, USA.
Small. 2015 Apr 17;11(15):1818-1828. doi: 10.1002/smll.201402564. Epub 2014 Dec 2.
A novel high-throughput magnetic tweezers-based 3D microchannel electroporation system capable of transfecting 40 000 cells/cm(2) on a single chip for gene therapy, regenerative medicine, and intracellular detection of target mRNA for screening cellular heterogeneity is reported. A single cell or an ordered array of individual cells are remotely guided by programmable magnetic fields to poration sites with high (>90%) cell alignment efficiency to enable various transfection reagents to be delivered simultaneously into the cells. The present technique, in contrast to the conventional vacuum-based approach, is significantly gentler on the cellular membrane yielding >90% cell viability and, moreover, allows transfected cells to be transported for further analysis. Illustrating the versatility of the system, the GATA2 molecular beacon is delivered into leukemia cells to detect the regulation level of the GATA2 gene that is associated with the initiation of leukemia. The uniform delivery and a sharp contrast of fluorescence intensity between GATA2 positive and negative cells demonstrate key aspects of the platform for gene transfer, screening and detection of targeted intracellular markers in living cells.
报道了一种基于新型高通量磁镊的3D微通道电穿孔系统,该系统能够在单个芯片上以40000个细胞/平方厘米的转染效率进行基因治疗、再生医学以及用于筛选细胞异质性的靶mRNA细胞内检测。单个细胞或单个细胞的有序阵列通过可编程磁场被远程引导至穿孔位点,细胞对齐效率高(>90%),从而使各种转染试剂能够同时递送至细胞中。与传统的基于真空的方法相比,本技术对细胞膜的损伤明显更小,细胞活力>90%,此外,还能使转染后的细胞被运输以便进一步分析。通过将GATA2分子信标递送至白血病细胞中以检测与白血病发生相关的GATA2基因的调控水平,展示了该系统的多功能性。GATA2阳性和阴性细胞之间荧光强度的均匀递送和鲜明对比证明了该平台在活细胞中进行基因转移、筛选和检测靶向细胞内标志物的关键特性。