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Cell electrotransfection device and its application

An electrotransfection and cell technology, which is applied in the field of cell transfection, can solve the problem that suspension cells cannot attach, and achieve good transfection effect, simple and easy-to-operate transfection process, and high application value

Active Publication Date: 2019-11-19
布莱迪杭州科学仪器有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Smaller suspension cells cannot attach to the surface of the nanotube substrate and cannot be transfected with this device

Method used

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  • Cell electrotransfection device and its application
  • Cell electrotransfection device and its application
  • Cell electrotransfection device and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Preparation of CuO nanowire electrode

[0061] In this embodiment, a CuO nanowire electrode is prepared by a thermal oxidation method, which specifically includes the following steps:

[0062] Purity greater than 99.9%, wire diameter 0.04mm, hole diameter 0.09mm, plain weave 200 mesh square hole copper mesh, punched into the required shape, and flattened with a tablet machine. Put the copper grid in 1.0mol·L -1 Soak in dilute hydrochloric acid solution for about 30 minutes to dissolve oxides and impurities on the surface. Rinse repeatedly with deionized water until the washing solution is neutral, and finally blow dry with nitrogen. Put the cleaned and dried copper grid into a quartz boat, put it into a temperature-programmed heating furnace, and raise the temperature to 500°C at a heating rate of 5°C / min in an atmosphere of 0.03MPa, and then heat at 500°C for 2 hours. , cooled to room temperature with the furnace. After the reaction, it can be seen that t...

Embodiment 2

[0063] Example 2 Construction of Cell Electrotransfection Device

[0064] The CuO nanowires prepared in Example 1 are used as electrodes (such as image 3 shown), the cell electrotransfection device was constructed. The electrotransfection device at least includes an electrotransfection control circuit, two CuO nanowire electrodes and an electrotransfection container. The electrotransfection control circuit can adopt existing technology, such as a signal generator (Puyuan DG1022U), which can produce accurate, stable, and low-distortion output signals, including five standard waveforms such as square wave, pulse, and sine wave. The arbitrary wave editing software Ultrawave can be used to output the required waveform, the output voltage range is 2mV~10V, and the frequency range is 1μHz~25MHz; the required waveform, voltage, pulse width, pulse number and other parameters can be flexibly adjusted according to the different electroporated cells. Two CuO nanowire electrodes are co...

Embodiment 3

[0067] Example 3 Cell electrotransfection device transfects adherent cells

[0068] Adherent cells were transfected by using the cell electrotransfection device constructed in Example 2 containing CuO nanowire electrodes. This example uses CHO cells as an example for illustration.

[0069] The transfection method comprises the following steps:

[0070] (1) The day before electroporation (electrotransfection), subculture the cells at a suitable density, so that the cells are in the logarithmic growth phase before transfection, and the confluence of the cell monolayer is about 70-80%;

[0071] (2) Soak two CuO nanowire electrodes, electrode holders and gaskets in 75% alcohol for 2 hours, then irradiate them under a UV lamp for 30 minutes and then use them. Place the CuO nanowires according to the method in Example 2 Electrodes and electrode holders;

[0072] (3) Add 1ml of DMEM medium (containing 10% serum, without antibiotics) to each well of a 6-well cell culture plate, and ...

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Abstract

The invention belongs to the technical field of cell electrotransfection and specifically relates to cell electrotransfection device and application thereof. The cell electrotransfection device at least comprises an electrotransfection control circuits, two electrodes and an electrotransfection container. The two electrodes are connected to a power supply in the electrotransfection control circuit. One of the two electrodes is used as a positive electrode, and the other electrode is used as a negative electrode. The two electrodes are arranged inside the electrotransfection container. One or two of the electrodes possess a nanowire structure on the surface thereof. The cell electrotransfection device of the invention can be used for electrotransfection of various cells. In addition, cells survival rate is high, and the electrotransfection effect is good. The cell electrotransfection device has high application value.

Description

technical field [0001] The invention belongs to the technical field of cell transfection, and in particular relates to a cell electrotransfection device and its application. Background technique [0002] Cell transfection is a specialized technique for introducing exogenous DNA, RNA, protein, polypeptide, small molecular compound, etc. into cells, and can be applied to eukaryotic cells and bacteria. With the continuous development of molecular biology and cell biology research, transfection has become a routine tool to study and control the gene function of cells. It is used more and more widely in biological experiments such as studying gene function, regulating gene expression, mutation analysis and protein production. [0003] Chimeric antigen receptor (CAR)-based cellular immunity is currently an emerging modality for the treatment of malignancies. CAR-modified T cells combine single-chain antibodies that recognize tumor-associated antigens with T-cell activation motif...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/42C12N15/87
CPCC12M35/02C12N15/87
Inventor 徐宇虹史家岑
Owner 布莱迪杭州科学仪器有限公司
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