Coplanar film electrodes facilitate bovine nuclear transfer cloning

Clow, Andrew L.; Gaynor, Paul; Oback, Björn

doi:10.1007/s10544-009-9302-z

Cited by 12 publications

(3 citation statements)

References 28 publications

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“…The cell electrofusion process was observed through the microscope and recorded by the CCD camera. The cell alignment, pairing, fusion efficiencies are calculated as the number of alignment cells, number of cell–cell twins × 2, and hybrid cells × 2 dividing the number of total cells, respectively []. Each experiment was repeated ten times.…”

Section: Methodsmentioning

confidence: 99%

A cell electrofusion microfluidic chip using discrete coplanar vertical sidewall microelectrodes

Yang

Qian

et al. 2012

Electrophoresis

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A novel cell electrofusion microfluidic chip using discrete coplanar vertical sidewall electrodes has been designed, fabricated, and tested. The device contains a serpentine-shaped microchannel with 22 500 pairs of vertical sidewall microelectrodes patterned on two opposing vertical sidewalls of the microchannel. The adjacent microelectrodes on each sidewall are separated by coplanar SiO(2) -Polysilicon-SiO(2) /silicon. This design of coplanar discrete vertical sidewall electrodes eliminates the "dead area" present in previous designs using continuous three-dimensional (3D) protruding sidewall electrodes, and generates uniform electric field along the height of the microchannel, leading to a lower voltage required for cell fusion compared to designs using 2D thin-film electrodes. This device is tested to fuse NIH3T3 cells under a low voltage (∼9 V). Almost 100% cells are aligned to the edge of the discrete microelectrodes, and cell-cell pairing efficiency reaches 70%. The electrofusion efficiency is above 40% of the total cells loaded into the device, which is much higher than traditional fusion methods and existing microfluidic devices using continuous 3D protruding sidewall microelectrodes.

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Section: Methodsmentioning

confidence: 99%

A cell electrofusion microfluidic chip using discrete coplanar vertical sidewall microelectrodes

Yang

Qian

et al. 2012

Electrophoresis

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“…The efficiencies of the cell pairing and electrofusion of this device are characterized by E p ¼ 2 Â paired cells/total cells and E f ¼ 2 Â fused cells/total cells, which are ensemble average of the 12 experiments obtained from the microscopic observation. 25,28 In each experiment, about 200 cells are observed within the microscope visual field.…”

Section: -5mentioning

confidence: 99%

A cell electrofusion microfluidic device integrated with 3D thin-film microelectrode arrays

Yang

Qian

et al. 2011

Biomicrofluidics

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A microfluidic device integrated with 3D thin film microelectrode arrays wrapped around serpentine-shaped microchannel walls has been designed, fabricated and tested for cell electrofusion. Each microelectrode array has 1015 discrete microelectrodes patterned on each side wall, and the adjacent microelectrodes are separated by coplanar dielectric channel wall. The device was tested to electrofuse K562 cells under a relatively low voltage. Under an AC electric field applied between the pair of the microelectrode arrays, cells are paired at the edge of each discrete microelectrode due to the induced positive dielectrophoresis. Subsequently, electric pulse signals are sequentially applied between the microelectrode arrays to induce electroporation and electrofusion. Compared to the design with thin film microelectrode arrays deposited at the bottom of the side walls, the 3D thin film microelectrode array could induce electroporation and electrofusion under a lower voltage. The staggered electrode arrays on opposing side walls induce inhomogeneous electric field distribution, which could avoid multi-cell fusion. The alignment and pairing efficiencies of K562 cells in this device were 99% and 70.7%, respectively. The electric pulse of low voltage ($9 V) could induce electrofusion of these cells, and the fusion efficiency was about 43.1% of total cells loaded into the device, which is much higher than that of the convectional and most existing microfluidics-based electrofusion devices.

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“…Regardless of method for cell fusion, higher fusion efficiency depends on perfect cell-cell contact instead of random cell pairing, as is the case in conventional methods. Controlled pairing of partner cell have been demonstrated using a high throughput cell pairing and fusion using chemical conjugation [ 45 ], field free micro structure assisted [ 8 , 31 ], electric field assisted cell pairing with better adoptability using hydrodynamic traps or constriction trapping [ 32 , 33 , 36 , 37 ]. Despite availability of various cell fusion platforms, cell fusion chip designing needs a deeper inspection towards performance parameters.…”

Section: Introductionmentioning

confidence: 99%

A microfluidic approach towards hybridoma generation for cancer immunotherapy

Pendharkar

et al. 2015

Oncotarget

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Dendritic cells/tumor fusions have shown to elicit anti-cancer immunity in different cancer types. However, the application of these vaccines for human cancer immunotherapy are limited by the instable quality and insufficient quanity of fusion cells. We present a cell electrofusion chip fabricated using soft lithography technique, which combines the rapid and precise cell pairing microstructures and the high yield electrofusion micro-electrodes to improve the cell fusion. The design uses hydrodynamic trapping in combination with positive dielectrophoretic force (pDEP) to achieve cell fusion. The chip consists of total 960 pairs of trapping channels, which are capable of pairing and fusing both homogeneous and heterogeneous types of cells. The fused cells can be easily taken out of the chip that makes this device a distinguishable from other designs. We observe pairing efficiency of 68% with fusion efficiency of 64%.

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Coplanar film electrodes facilitate bovine nuclear transfer cloning

Cited by 12 publications

References 28 publications

A cell electrofusion microfluidic chip using discrete coplanar vertical sidewall microelectrodes

A cell electrofusion microfluidic chip using discrete coplanar vertical sidewall microelectrodes

A cell electrofusion microfluidic device integrated with 3D thin-film microelectrode arrays

A microfluidic approach towards hybridoma generation for cancer immunotherapy

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