Preparation method of silver nanowire array and application of silver nanowire array in plasmid transformation of stem cells

A nanowire array and stem cytoplasm technology, which is applied in the preparation of silver nanowire arrays and its application in stem cell plasmid transformation, can solve the problems of low overall efficiency and different cell transfection efficiency, and achieves simple operation and reduced table. The influence of epigenetics, the effect of solving low transformation efficiency

Inactive Publication Date: 2018-06-01
章毅 +7
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Cell transfection efficiencies vary from tissue and species origin and are generally not high

Method used

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  • Preparation method of silver nanowire array and application of silver nanowire array in plasmid transformation of stem cells
  • Preparation method of silver nanowire array and application of silver nanowire array in plasmid transformation of stem cells

Examples

Experimental program
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Effect test

Embodiment 1

[0022] With a porous alumina plate as the cathode and a carbon rod as the anode, the H 2 SO 4 / AgNO 3 The mixed solution is an electrolyte, the AC voltage is about 12 volts, and the electrodeposition is at room temperature for 29 minutes. During the deposition process, the magnetron keeps stirring the solution to avoid local overheating of the solution. Afterwards, the porous alumina template is rinsed with deionized water to remove H on template 2 SO 4 solution.

[0023] The porous alumina template deposited with silver nanowires is put into 0.2 mol / L sodium hydroxide solution to dissolve the aluminum base, and prepare the silver nanowire array with the substrate.

Embodiment 2

[0025] The silver nanowire array was punched into a 33mm diameter disc and autoclaved and autoclaved, and put into a 35mm diameter cell culture dish ( figure 1 ), prepared as a nanowire array delivery device. The DMEM medium containing 7721 liver cancer cells was added to the device, and the expression plasmid of green fluorescent protein was added dropwise. After the cells were cultured for 3 days, 90% of the cells produced green fluorescence under the detection of an inverted fluorescent microscope, which indicated that the expression plasmid containing the green fluorescent protein had been transported into the cells and was successfully expressed with biological activity. Cell viability was also maintained at 90%.

Embodiment 3

[0027] The silver nanowire array was punched into a 32mm disc, and put into a commonly used 6-well cell culture plate to prepare a nanowire array delivery device (see figure 2 ). 2ml of DMEM medium (10% fetal bovine serum (GIBICO), 5% horse serum (GIBICO)) containing mouse neural stem cells C17.2 was added to the device, and the expression plasmid pEGFP-C1 of green fluorescent protein was added dropwise. Neural After the stem cells have been cultured for 3 to 5 days, 90% of the living cells have green fluorescence under the detection of an inverted fluorescent microscope, which indicates that the expression plasmid containing green fluorescent protein has been transported into the cells and has biological activity. Neural The survival rate of stem cell C17.2 also maintained at 80%-90%.

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Abstract

The invention relates to application of a nanowire array in preparation of a conveying device for converting plasmid DNA of stem cells; the nanowire array is capable of absorbing plasmid and enteringthe stem cells so as to realize plasmid delivery. According to the application, the ratio of cells to plasmids does not need to be strictly controlled; the time for incubating mammalian cells and plasmid DNA together is generally 2-3 days, and a conversion rate of 80-90% is basically reached. In the case of slow cell growth, the conversion rate is facilitated to be increased by prolonging the incubation time; especially, a full consideration needs to be taken by combining actual conditions during stem cell culture; the time for incubating the stem cells and the plasmid DNA together is generally 3-5 days.

Description

technical field [0001] The invention relates to the application of the silver nanowire array, in particular to the application of the silver nanowire array in the preparation of mammalian cells and various mammalian stem cell plasmid DNA delivery devices. Background technique [0002] Transducing plasmids into mammalian cells is an important step in gene expression analysis and cell engineering. At present, the commonly used methods for introducing plasmids into mammalian cells are mainly divided into two categories: one is electroporation. The method adopts an electroporation instrument and uses a pulsed field current to instantaneously introduce a plasmid into a cell through a small hole in the cell membrane. Manufacturers of cell electroporation instruments mainly include American BioRad, American BTX, American Invitrogen, and German Eppendorf. The electroporation method not only needs to strictly control the conditions, such as the voltage intensity, current intensity ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/87C12N15/85C25D3/46B82Y30/00B82Y40/00B82Y5/00
CPCB82Y5/00B82Y30/00B82Y40/00C12N15/85C12N15/87C12N2800/107C25D3/46
Inventor 章毅王艺翟晓文周渊峰陈亮陆薇黄海
Owner 章毅
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