Flow-type electroporation device

An electrotransfection and flow-through technology, applied in the field of electrotransfection, can solve the problems of difficult to control the processing time of cells, small volume of cell fluid processing, difficult to quickly process a large number of samples, etc. The effect of high electrotransfection efficiency

Pending Publication Date: 2020-03-10
ETTA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On this basis, the researchers invented the parallel connection of multiple electrotransfection chambers. This technology has its advantages, but it cannot fundamentally solve the difficulty of quickly processing a large number of samples.
[0010] The technical problem to be solved by the pr

Method used

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  • Flow-type electroporation device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The flow electrotransfection device provided by the present invention is as Figure 1-3 As shown, it includes an insulating tube body (1), a first electrode (2), a second electrode (3), a bracket (5) for fixing the first electrode, a bracket (6) for fixing the second electrode, and a liquid inlet of the tube body (7), the liquid outlet of the pipe body (8), the pipe wall of the opening at both ends of the insulating pipe body (1) is provided with a groove, the end face of the groove is connected with the bracket (5) for fixing the first electrode, and the second electrode is fixed. The ring-shaped end faces of the brackets (6) of the electrodes are of the same size, the brackets (5) for fixing the first electrode and the brackets (6) for fixing the second electrode are respectively fixed at the grooves at both ends by gluing, and the liquid inlet of the pipe body ( 7) The size of the end face and the end face of the liquid outlet (8) of the pipe body is the same as that...

Embodiment 2

[0032] Collect 293F cells (human embryonic kidney cells, suspension type) in the logarithmic growth phase, rotate at 1000 rpm, centrifuge for 5 minutes, discard the supernatant, and resuspend with 10 ml of commercially available EBEL electroporation buffer cells such that the cell density is 2.0 x 10 7 pcDNA3.1-GFP, which needs to be electrotransfected into the cells, was added to make the concentration of the plasmid 40 μg / ml, and mixed gently.

[0033]In the flow electroporation device prepared in the present invention, the cell suspension mixed with 293F cells and plasmid pcDNA3.1-GFP is pumped into the tube body through the annular gap at the inlet port through a complete set of peristaltic pump plus hose device, Then it flows out of the pipe body through the annular gap at the outlet end.

[0034] Used in conjunction with flow electrotransfection equipment, electrical stimulation was performed under the following conditions: flow rate 5.5 ml / min, voltage 180 volts, pulse...

Embodiment 3

[0036] Using the flow-type electrotransfection device of the present invention and the non-flow-type electrotransfection device, 293F cells were used to conduct a biological assay experiment comparison, and the non-flow-type electrotransfection device was an electrode cup routinely used in the field.

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Abstract

The invention provides a flow-type electroporation device. The device includes a pipe body, a first electrode, a second electrode, a support for fixing the first electrode, and a support for fixing the second electrode; and the device is characterized in that pipe openings of two ends of the pipe body are provided with grooves, the support for fixing the first electrode and the support for fixingthe second electrode are respectively arranged in the grooves, the shapes of the first electrode and the second electrode are the same as the shape of the cross section of the middle part of the pipebody, the pipe body forms a cavity with the first electrode and the second electrode, a gap is arranged between the support for fixing the first electrode and the support for fixing the second electrode, and a liquid enters and/or flows out of the cavity through the gap. The flow-type electroporation device provided by the invention can form an electroporation only when the cavity is filled with the liquid, ensures the same time for the cell electroporation, and improves the cell processing capacity while improving the electroporation efficiency.

Description

technical field [0001] The invention relates to the field of electrotransfection, relates to a flow electrotransfection device, in particular to a method for performing electrotransfection only after the device is filled with fluid. Background technique [0002] The cell membrane is a layer of thin film surrounding the periphery of the cell, which is a permeability barrier for the selective material exchange between the cell and the outside world. The cell membrane makes the cell an independent living unit and has a relatively stable internal environment. Some substances in the surrounding environment can pass through the cell membrane, while others cannot. Cells can absorb nutrients from the surrounding environment through the cell membrane, discharge metabolites, and make the transport of substances reach a state of balance. Therefore, the basic function of the cell membrane is to maintain the relative stability of the intracellular microenvironment and selectively excha...

Claims

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Application Information

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IPC IPC(8): C12M1/42
CPCC12M35/02
Inventor 戴晓兵朱士英徐莉
Owner ETTA BIOTECH
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