Cell electrotransfection device and application thereof

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: 2017-08-01
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 application thereof
  • Cell electrotransfection device and application thereof
  • Cell electrotransfection device and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Preparation of CuO nanowire electrode

[0061] In this embodiment, the thermal oxidation method is used to prepare CuO nanowire electrodes, which specifically includes the following steps:

[0062] A 200-mesh square-hole copper net with a purity of greater than 99.9%, a wire diameter of 0.04mm, a hole diameter of 0.09mm, and a plain weave is punched into the desired shape and flattened with a tablet press. Place the copper mesh at 1.0mol·L -1 Soak in the diluted hydrochloric acid solution for about 30 minutes to dissolve the oxides and impurities on the surface. Then rinse with deionized water repeatedly until the lotion is neutral, and finally, blow dry with nitrogen. Put the cleaned and dried copper mesh into a quartz boat and send it to a temperature-programmed heating furnace. In an atmosphere of 0.03MPa, the temperature is raised to 500°C at a heating rate of 5°C / min, and then heated at 500°C for 2h , Cool down to room temperature with the furnace. After th...

Embodiment 2

[0063] Example 2 Construction of cell electrotransfection device

[0064] The CuO nanowires prepared in Example 1 were used as electrodes (e.g. image 3 Shown) to construct a cell electrotransfection device. The electrotransfection device includes at least one electrotransfection control circuit, two CuO nanowire electrodes and an electrotransfection container. The electrotransfection control circuit can use existing technology, such as a signal generator (Puyuan DG1022U), which can generate accurate, stable, and low-distortion output signals, including five standard waveforms such as square wave, pulse, and sine wave. The arbitrary waveform editing software Ultrawave can be used to output the required waveforms. 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 electroporation cells. The two CuO nanowire electrodes are conn...

Embodiment 3

[0067] Example 3 Transfection of adherent cells with cell electrotransfection device

[0068] The cell electrotransfection device containing CuO nanowire electrodes constructed in Example 2 was used to transfect adherent cells. This example uses CHO cells as an example.

[0069] The transfection method includes the following steps:

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

[0071] (2) Place the two CuO nanowire electrodes, electrode holders and gaskets in 75% alcohol for 2 hours, and then irradiate them under a UV lamp for 30 minutes before use. Place the CuO nanowires in the same way as in Example 2. Electrode and electrode holder;

[0072] (3) Add 1ml DMEM medium (containing 10% serum, without antibiotics) to each well of the 6-well cell culture plate, and place it in CO at 37°C. 2 Preheat in t...

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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 specifically 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 molecule compounds, 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 for studying and controlling cell gene functions. It has become more and more widely used in biological experiments such as studying gene function, regulating gene expression, mutation analysis and protein production. [0003] Cellular immunity based on chimeric antigen receptor (CAR) has become an emerging form of treatment of malignant tumors. CAR modified T cell is a combination of single-chain antibody that recognizes tumor-associated antigen and T cell activation ...

Claims

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Application Information

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Patent Type & Authority Applications(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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