Gene Transfection of Mammalian Cells Using Membrane Sandwich Electroporation

Fei, Zhengzheng; Wang, Shengnian; Xie, Yizhen; Henslee, Brian E; Koh, Chee Guan; Lee, L James

doi:10.1021/ac070482y

Cited by 60 publications

(80 citation statements)

References 13 publications

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“…The permeabilization area can be controlled with the pulse amplitude and the degree of permeabilization can be controlled with the duration of pulses, numbers of pulses, where longer pulses provide a larger perturbation area in the cell membrane [34,35]. In earlier studies of micro/nanofluidic based single cell electroporation, authors analyze cellular content and cellular properties [36][37][38][39], transfection of cells [17,[40][41][42] and inactivating cells [43][44][45] with the use of micro-channel based electroporation [46][47][48][49], micro-capillary based electroporation [50][51][52], electroporation with solid microelectrode [36,[53][54][55], membrane sandwich based microfluidic electroporation [56,57], microarray single cell electroporation [58], optofluidic based microfluidic devices [59][60][61][62][63][64][65], etc. Table 1 describes in detail micro/nanofluidic based single cell transfection, cell lysis, cell type with species, potential difference, pulse duration, etc.…”

Section: Micro/nanofluidic Devices For Single Cell Electroporationmentioning

confidence: 99%

Recent Trends on Micro/Nanofluidic Single Cell Electroporation

Santra

Tseng

2013

Micromachines

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Abstract:The behaviors of cell to cell or cell to environment with their organelles and their intracellular physical or biochemical effects are still not fully understood. Analyzing millions of cells together cannot provide detailed information, such as cell proliferation, differentiation or different responses to external stimuli and intracellular reaction. Thus, single cell level research is becoming a pioneering research area that unveils the interaction details in high temporal and spatial resolution among cells. To analyze the cellular function, single cell electroporation can be conducted by employing a miniaturized device, whose dimension should be similar to that of a single cell. Micro/nanofluidic devices can fulfill this requirement for single cell electroporation. This device is not only useful for cell lysis, cell to cell fusion or separation, insertion of drug, DNA and antibodies inside single cell, but also it can control biochemical, electrical and mechanical parameters using electroporation technique. This device provides better performance such as high transfection efficiency, high cell viability, lower Joule heating effect, less sample contamination, lower toxicity during electroporation experiment when compared to bulk electroporation process. In addition, single organelles within a cell can be analyzed selectively by reducing the electrode size and gap at nanoscale level. This advanced technique can deliver (in/out) biomolecules precisely through a small membrane area (micro to nanoscale area) of the single cell, known as localized single cell membrane electroporation (LSCMEP). These articles emphasize the recent progress in micro/nanofluidic single cell electroporation, which is potentially beneficial for OPEN ACCESSMicromachines 2013, 4 334 high-efficient therapeutic and delivery applications or understanding cell to cell interaction.

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Section: Micro/nanofluidic Devices For Single Cell Electroporationmentioning

confidence: 99%

Recent Trends on Micro/Nanofluidic Single Cell Electroporation

Santra

Tseng

2013

Micromachines

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“…Recently, electroporation was integrated into microfluidic devices by using various types of mechanisms [13][14][15][16][17] . In comparison to conventional electroporation methods, microfluidic systems have some advantages, including the use of a lower applied electric field, lower sample and reagent consumption and reduced Joule heating 6 .…”

Section: Introductionmentioning

confidence: 99%

Continuous medium exchange and optically induced electroporation of cells in an integrated microfluidic system

et al. 2015

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Optically induced electroporation (OIE) is a promising microfluidic-based approach for the electroporation of cell membranes. However, previously proposed microfluidic cell-electroporation devices required tedious sample pre-treatment steps, specifically, periodic media exchange. To enable the use of this OIE process in a practical protocol, we developed a new design for a microfluidic device that can perform continuous OIE; i.e., it is capable of automatically replacing the culture medium with electroporation buffers. Integrating medium exchanges on-chip with OIE minimises critical issues such as cell loss and damage, both of which are common in traditional, centrifuge-based approaches. Most importantly, our new system is suitable for handling small or rare cell populations. Two medium exchange modules, including a micropost array railing structure and a deterministic lateral displacement structure, were first adopted and optimised for medium exchange and then integrated with the OIE module. The efficacy of these integrated microfluidic systems was demonstrated by transfecting an enhanced green fluorescent protein (EGFP) plasmid into human embryonic kidney 293T cells, with an efficiency of 8.3%. This result is the highest efficiency reported for any existing OIE-based microfluidic system. In addition, successful co-transfections of three distinct plasmids (EGFP, DsRed and ECFP) into cells were successfully achieved. Hence, we demonstrated that this system is capable of automatically performing multiple gene transfections into mammalian cells.

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“…A membrane sandwich electroporation device has been developed by integrating nanoporous membrane with an electrokinetic microfluidic platform. 4 We have accomplished tests with dextran (as model drugs) and reporting genes (GFP and SeAP) delivered into NIH 3T3 fibroblast cells. The efficiency is much better than the existing electroporation method.…”

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confidence: 99%

Overview of polymer micro/nanomanufacturing for biomedical applications

Farson

et al. 2008

Adv Polym Technol

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Micro/nanotechnology is initiated from the electronics industry.In recent years, it has been extended to micro/nanoelectromechanic system for producing miniature devices based on silicon and semiconductor materials. However, the use of these hard materials alone is inappropriate for many biomedical devices. Soft polymeric materials possess many attractive properties such as high toughness and recyclability. Some possess excellent biocompatibility, are biodegradable, and can provide various biofunctionalities. Proper combinations of micro/nanoelectronics, polymers, and biomolecules can lead to new and affordable medical devices. In this paper, we briefly review several cleanroom and noncleanroom techniques related to micro/nanomanufacturing of polymeric materials.

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Gene Transfection of Mammalian Cells Using Membrane Sandwich Electroporation

Cited by 60 publications

References 13 publications

Recent Trends on Micro/Nanofluidic Single Cell Electroporation

Recent Trends on Micro/Nanofluidic Single Cell Electroporation

Continuous medium exchange and optically induced electroporation of cells in an integrated microfluidic system

Overview of polymer micro/nanomanufacturing for biomedical applications

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