CN110885857B - Intermittent flow type formula electrotransfection device - Google Patents
Intermittent flow type formula electrotransfection device Download PDFInfo
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- CN110885857B CN110885857B CN201811054671.0A CN201811054671A CN110885857B CN 110885857 B CN110885857 B CN 110885857B CN 201811054671 A CN201811054671 A CN 201811054671A CN 110885857 B CN110885857 B CN 110885857B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
Abstract
The invention discloses an intermittent flow type electrotransfection device, which comprises a tube body, a first electrode and a second electrode, and is characterized in that the first electrode and the second electrode are parallel at two ends of the tube body, and a cavity for accommodating a target liquid sample is formed inside the tube body and the first electrode and the second electrode; the pipe body is provided with at least one liquid inlet and outlet and at least one liquid inlet and outlet valve, and the liquid inlet and outlet valve is positioned at the liquid inlet and outlet of the pipe body and controls the opening and closing of the liquid inlet and outlet; the first electrode and/or the second electrode are relatively movable along the tube. The intermittent flow type electrotransfection device of the invention utilizes pressure to avoid bubbles generated in the electrotransfection process, improves electrotransfection efficiency, and realizes rapid treatment of a large number of samples.
Description
Technical Field
The invention relates to the field of electrotransfection, in particular to an intermittent flow type electrotransfection device, which is capable of inhibiting bubbles generated by electrotransfection in a mode of electrode moving and pressurizing.
Background
The cell membrane is a membrane surrounding the periphery of the cell and is a permeable barrier for selective substance exchange between the cell and the outside. The cell membrane makes the cell an independent unit of life and has a relatively stable internal environment. Some substances in the surrounding environment may pass through the cell membrane while others may not. Cells can take up nutrients from the surrounding environment through the cell membrane and discharge metabolites, so that the transport of substances reaches an equilibrium state. The basic function of the cell membrane is therefore to maintain a relatively stable intracellular microenvironment and to selectively exchange substances with the external environment.
It has been found that if a certain intensity of electrical stimulation is applied to cells for a certain period of time, some micropores are induced in the cell membrane, so that the permeability of the cells is enhanced, and that cell Electroporation (electric corporation) refers to the biophysical process of forming transient micropores in the lipid bilayer of the cell membrane under the action of an applied pulsed electric field. Electrotransfection (Electrotransfection) is a technique that uses electroporation to introduce exogenous biological macromolecules, such as DNA, RNA, or proteins, into cells. When the cell membrane is electroporated, its permeability and membrane conductance are transiently increased, allowing molecules that normally cannot pass through the cell membrane, such as hydrophilic molecules, DNA, proteins, viral particles, drug particles, etc., to enter the cell through the microwells. After the electrical stimulation is removed in a short period of time, the micropores on the cell membrane disappear and the cell membrane becomes a selectively permeable barrier again. Compared with the traditional chemical transfection and virus transfection, the electrotransfection has the advantages of no chemical pollution, no permanent damage to cells, higher efficiency and the like, and has wide application prospect in the fields of biology, molecular biology, clinical medicine and the like.
Although the mechanism of electrotransfection is not entirely clear, cell electrotransfection is well known herein, including movement of the membrane lipid bilayer, resulting in the formation of temporary micropores in the membrane, allowing exogenous molecules to enter the cell through the micropores.
In the prior art, there are three main types of methods for completing the cell electrotransfection process:
cells are placed between a pair of parallel electrodes spaced apart by a few millimeters to a few centimeters. The cells are electrically stimulated in the electric field between the electrodes to achieve the purpose of electrotransfection. For example, US patent US5389069.
The miniature needle-shaped electrode is used for being pricked into tissue or cell fluid to perform electric shock on cells, so that the purpose of electric transfection is achieved. For example, US patent US5389069.
A chamber is placed between a pair of parallel electrodes such that a suspension of cells is shocked while flowing in the chamber. For example, US patent US6773669.
Chinese patent CN201010242144 discloses a flow electrotransfection device and system, the system comprising: a flow electrotransfection device comprising: the electrode is arranged in parallel and in pairs, and each pair of electrodes comprises an anode and a cathode which are oppositely arranged; a fluid flow restricting channel disposed over the electrode; the channel starting end is provided with a plurality of inlet branch channels and is converged into a main channel, the ending end is provided with a plurality of outlet branch channels, and a top cover with a plurality of fluid inlets and outlets is arranged above the channels; a syringe pump connected by tubing to the inlet and outlet of the top cover in the flow electrotransfection device to control the flow rate of the fluid; and the voltage source is connected with the electrode through the electric connecting piece and is used for setting and generating pulse voltage. The flow type electrotransfection system utilizes the fluid channel and the connected injection pump to realize continuous flow of various suspensions in the fluid channel, so that the process of cell electrotransfection can be continuously carried out, and a large amount of samples can be rapidly processed.
Chinese patent CN201610806987 discloses a disposable for electrotransfection of cells comprising: a fluid compartment inside the disposable; a first fluid port for providing a cell suspension to the fluid compartment; and a second fluid port for delivering a fluid comprising at least one compound to be electrotransfected into the cell to the fluid compartment; a first electrode and a second electrode disposed in the fluid compartment; at least one outlet port delivering the fluid from the fluid compartment, wherein the first and second fluid ports are in fluid communication with a mixing channel in fluid communication with the fluid compartment.
However, the flow type electrotransfection device disclosed above can generate a large amount of bubbles in the actual electrotransfection process, and the generated bubbles adhere to the electrode plates to influence the uniformity of the electric field, so that the electrotransfection effect is unstable. The inventor finds through search that the technical problem of generating bubbles in the electrotransfection process is not solved in the field of flow electrotransfection.
Disclosure of Invention
The invention aims at the technical problems in the prior art and provides an intermittent flow type electrotransfection device for inhibiting bubbles in the electrotransfection process by a movable electrode pressurizing mode. It is another object of the invention to improve the stability of electrotransfection devices and to improve the efficiency of electrotransfection.
In order to achieve the above purpose, the technical scheme of the intermittent flow type electrotransfection device provided by the invention is summarized as follows: the intermittent flow type electrotransfection device comprises a tube body, a first electrode and a second electrode, and is characterized in that the first electrode and the second electrode are parallel at two ends of the tube body, and a cavity for accommodating a target liquid sample is formed inside the tube body and the first electrode and the second electrode; the pipe body is provided with at least one liquid inlet and outlet and at least one liquid inlet and outlet valve, and the liquid inlet and outlet valve is positioned at the liquid inlet and outlet of the pipe body and controls the opening and closing of the liquid inlet and outlet; the first electrode and/or the second electrode are relatively movable along the tube.
Preferably, the pipe body is provided with a vent pipe, the vent pipe is provided with a vent valve, further preferably, the joint of the vent pipe and the pipe body is provided with a vent valve, the vent pipe is positioned between the first electrode and the second electrode, and the tail end of the vent pipe is provided with an air filtering device.
Preferably, the pipe body is provided with at least two liquid inlets and outlets, and the liquid inlets and outlets can be liquid inlets and liquid outlets, and at least two liquid inlet and outlet valves are correspondingly arranged; the liquid inlet and outlet valve can be a liquid sample injection one-way valve or a liquid sample outlet one-way valve; it is further preferable that the tube body is provided with at least one liquid inlet and one liquid outlet, at least one liquid sampling check valve and one liquid sampling check valve;
preferably, the cross sections of the first electrode and the second electrode are the same as or close to the cross section of the inner part of the tube body, more preferably, the cross section of the inner part of the tube body is square, and the cross section of the inner part of the tube body can be round or other shapes;
preferably, sealing devices are arranged at the contact positions of the first electrode and the second electrode and the pipe body, and the sealing devices are preferably sealing gaskets, sealing rubber strips or other devices with sealing performance;
preferably, when the liquid inlet and outlet valve and the ventilation valve are closed, the tube body, the first electrode and the second electrode form a closed cavity;
preferably, when the liquid inlet and outlet valve and the ventilation valve are closed, the pressure value in the closed cavity is 1.0-2.5Mpa after the first electrode and/or the second electrode move relative to the pipe body; it is further preferred that the pressure value inside the cavity is 1.1-2.0Mpa; it is further preferred that the pressure value inside the cavity is 1.2-1.5Mpa;
preferably, the device further comprises a plurality of guide pipes, which are divided into a liquid inlet pipe and a liquid outlet pipe, and the guide pipes and the pipe body are arranged in a sealing way;
preferably, the device further comprises a pump for flowing liquid out of or into the cavity, or for exhausting or filling gas within the cavity, the type of pump including, but not limited to, peristaltic pump, air pump, liquid pump, syringe pump, etc.;
preferably, the device further comprises a pressurizing device enabling the first electrode and/or the second electrode to relatively move along the tube body, wherein the pressurizing device comprises a balloon, a hydraulic rod, a pneumatic rod, a spiral pressurizing device, an electromagnetic force pressurizing device and the like;
preferably, the first electrode and/or the second electrode are provided with an operating position and a non-operating position, and the electrodes are moved from the non-operating position to the operating position and can provide pressure for the interior of the cavity; it is further preferred that the liquid inlet and outlet is located between the first electrode and the second electrode. It is further preferred that the liquid inlet and outlet is located between an operative position and an inoperative position in which the first electrode and/or the second electrode is arranged.
Preferably, the second electrode is fixedly arranged at an orifice at one end of the tube body.
The invention relates to a preparation method of a device, which comprises the steps of firstly preparing a pipe body, selecting plastic capable of being cured after heating as a material of the pipe body, preparing the pipe body by utilizing a mould for preparing the pipe body, respectively arranging a liquid sample inlet and a liquid sample outlet on the pipe body, respectively arranging a relatively parallel electrode at two ends of the pipe body, and injecting two metal sheets with the same shape and size into a plastic frame to form a first electrode and a second electrode. And one side of the electrode, which is not contacted with the cavity, is connected with the pressurizing device, and soft adhesive tapes are arranged around the electrode plastic frame. Fixedly mounting the second electrode, and connecting the first electrode with the pressurizing device so that the first electrode can displace under the pressure generated by the pressurizing device; or the first electrode and the second electrode are respectively connected with the pressurizing device, so that the first electrode and the second electrode can displace under the pressure generated by the pressurizing device.
The technical scheme of the invention has the beneficial effects that:
the intermittent flow type electrotransfection device of the invention can process a large amount of liquid samples.
The intermittent flow type electric transfection device pressurizes liquid in the electric transfection process, inhibits air bubbles from generating, reduces electric transfection efficiency fluctuation caused by air bubbles adhesion, and improves the stability of the electric transfection device.
The intermittent flow type electric transfection device has novel structural design and originality.
Drawings
FIG. 1 is a schematic diagram of an exemplary batch flow electrotransfection device of the present invention;
FIG. 2 is a schematic diagram of an exemplary batch flow electrotransfection device of the present invention;
the accompanying drawings annotate: the device comprises a first electrode, a second electrode, a 3-tube body, a 4-liquid inlet tube, a 5-liquid outlet tube, a 6-pressurizing device, a 7-first electrode non-working position, a 8-first electrode working position, a 9-second electrode non-working position, a 10-second electrode working position, a 11-sample-injection one-way valve, a 12-sample-outlet one-way valve, a 13-vent tube and a 14-vent valve.
Description of the embodiments
Example 1
Integral structural design 1
The whole structure of the intermittent flow type electrotransfection device provided by the invention is shown in figure 1, the pipe orifices at the two ends of the pipe body (3) are sealed, wherein one end of the pipe orifice adopts the second electrode (2) as the sealing arrangement, the other end of the pipe orifice adopts the first electrode (1) as the sealing arrangement, the pipe body (3) is provided with a vent pipe (13), and the joint of the vent pipe (13) and the pipe body (3) is provided with a vent valve (14). The first electrode (1) is provided with a working position (8) and a non-working position (7) in the pipe body (3), the first electrode (1) is connected with the pressurizing device (6), the pressurizing device (6) is preferably a hydraulic rod, and the pressurizing device (6) can push the first electrode (1) to move from the non-working position (7) to the working position (8), so that liquid borne in the cavity bears certain pressure. An electrical pulse is applied to the first electrode (1) and the second electrode (2), the electrical pulse causing an electric field to form within the cavity such that the cell membrane forms a microwell, thereby enabling a target substance within the liquid sample to enter the cell. After the electrotransfection is completed, the pressurizing means (6) are able to pull the first electrode (1) back from the working position (8) to the non-working position (7). The above operation was repeated for intermittent flow electrotransfection.
Example two
Integral structural design 2
The whole structure of the intermittent flow type electrotransfection device provided by the invention is shown in fig. 2, the end caps at the two end orifices of the tube body (3) are respectively arranged in a sealing way by adopting a first electrode (1) and a second electrode (2), wherein the first electrode (1) and the second electrode (2) are connected with a pressurizing device (6), and the pressurizing device (6) is preferably a hydraulic rod. The pipe body (3) is provided with a vent pipe (13), and a vent valve (14) is arranged at the joint of the vent pipe (13) and the pipe body (3). The first electrode (1) is provided with an operating position (8) and a non-operating position (7) in the tube body (3), and the second electrode (2) is provided with an operating position (10) and a non-operating position (9) in the tube body. Depending on the relative movement of the first electrode (1) and the second electrode (2), the following three operating states can be set.
In the first working state, the pressurizing device (6) at the pipe orifices at the two ends pushes the first electrode (1) and the second electrode (2) to move from the non-working positions (7) and (9) to the working positions (8) and (10), so that the liquid in the cavity bears a certain pressure, at the moment, electric pulses are applied to the first electrode (1) and the second electrode (2), and an electric field is formed in the cavity by the electric pulses, so that micropores are formed in a cell membrane, and target substances in a liquid sample can enter the cell. After the electrotransfection is completed, the pressurizing device (6) can pull the first electrode (1) and the second electrode (2) back from the working position to the non-working position.
In the second working state, the second electrode (2) is fixed at the non-working position (9), the pressurizing device (6) pushes the first electrode (1) to move from the non-working position (7) to the working position (8) so that the liquid in the cavity bears a certain pressure, at the moment, electric pulses are applied to the first electrode (1) and the second electrode (2), and an electric field is formed in the cavity by the electric pulses, so that micropores are formed in a cell membrane, and a target substance in a liquid sample can enter the cell. After the electrotransfection is completed, the pressurizing means (6) are able to pull the first electrode (1) back from the working position (8) to the non-working position (7).
In the third working state, the first electrode (1) is fixed at the non-working position (7), the pressurizing device (6) pushes the second electrode (2) to move from the non-working position (9) to the working position (10) so that the liquid in the cavity bears a certain pressure, at the moment, electric pulses are applied to the first electrode (1) and the second electrode (2), and an electric field is formed in the cavity by the electric pulses, so that a cell membrane has permeability, and a target substance in a liquid sample can enter the cell. After the electrotransfection is completed, the pressurizing means (6) are able to pull the second electrode (2) back from the working position (10) to the non-working position (9).
Example III
Intermittent liquid feeding mode
When liquid is fed, a sample injection one-way valve (11) is opened, a sample discharge one-way valve (12) is closed, a ventilation valve (14) is opened, the gas with the same volume as the pipe body (3) is pumped out by an air pump through a ventilation pipe (13), and a target liquid sample enters the cavity from a liquid inlet pipe (4); or the air pump is not used for pumping out the air with the same volume as the pipe body (3) through the vent pipe (13), and the peristaltic pump is used for pressing the target liquid sample into the cavity from the liquid inlet pipe (4); after the cavity is full, the sampling check valve (11) and the ventilation valve (14) are closed.
After the electrotransfection is finished, a sample outlet one-way valve (12) and a ventilation valve (14) are opened, a gas pump is used for filling the gas with the same volume as the cavity into the pipe body through the ventilation pipe (14), and a target liquid sample is discharged from a liquid outlet pipe (5); or the air pump is not used for filling the air with the volume equal to that of the tube body into the cavity through the vent tube (14), and the target liquid sample is discharged from the liquid outlet tube (5) by gravity; after the target liquid sample is completely discharged, the sample outlet check valve (12) is closed, the sample inlet check valve (11) is opened, and the above liquid inlet actions are repeated.
Example IV
Method for pressurizing liquid in cavity
After the target liquid enters and fills the cavity through the sampling check valve (11), the sampling check valve (11) is closed, and at the moment, a specific pressure is applied to the first electrode (1) and/or the second electrode (2) through the pressurizing device (6) so that the first electrode and/or the second electrode move from the non-working positions (7) (9) to the working positions (8) (10), the pressurizing of the liquid is finished, and the electric transfection of the liquid is carried out in a pressurized state.
Example five
Biological assay 1
Collecting CHO-S cells (Chinese hamster ovary cells) in logarithmic growth phase, centrifuging at 1000 rpm for 5 min, discarding supernatant, and resuspending the cells with electrotransfection buffer to give a cell density of 1X10 7 Plasmid pCDNA3.1-GFP, which requires electrotransfection into cells, was added at a volume of each ml to allow plasmid to passThe concentration was 20ug/ml and gently mixed well. The intermittent flow type electrotransfection device is shown in figure 1, the end caps at the two ends of the tube body are respectively sealed by a first electrode and a second electrode, and the first electrode is connected with a hydraulic rod. Before the liquid enters the cavity, a sample injection one-way valve is opened, a sample discharge one-way valve is closed, a ventilation valve is opened, the gas with the same volume as the pipe body is pumped out by an air pump through a ventilation pipe, and a target liquid sample enters the cavity from a liquid inlet pipe; after the cavity is full, the sampling check valve and the ventilation valve are closed. The first electrode is maintained in the inactive position and the liquid is not pressurized. At this time, an electric pulse is applied to the first electrode and the second electrode, the pulse voltage is 180 volts, the pulse width is 6 milliseconds, the pulse times are 2 times, the pulse interval is 1 second, the electric pulse enables an electric field to be formed in the cavity, so that a cell membrane forms micropores, and target substances in the liquid sample can enter the cells. After the electrotransfection is finished, the sample outlet one-way valve and the ventilation valve are opened, the air pump is used for filling the air with the volume equal to that of the cavity body through the ventilation pipe, and the target liquid sample is discharged from the liquid outlet pipe. The transfected cell suspension was placed in a centrifuge tube, 1000 rpm, and centrifuged for 5 minutes. Removing supernatant, adding CD-OptiCHO culture medium to resuspend cells, inoculating, and culturing in triangular conical flask with culture density of 2×10 6 And/ml, then placing on a shaking table for culture, wherein the rotation speed of the shaking table is 125rpm/min, and the culture conditions are as follows: the temperature was 37℃and the carbon dioxide concentration was 5%. After 24 hours, the cells were observed under a fluorescence microscope and examined for electrotransfection efficiency and cell viability using a flow cytometer.
Example six
Bioassay 2
Collecting CHO-S cells (Chinese hamster ovary cells) in logarithmic growth phase, centrifuging at 1000 rpm for 5 min, discarding supernatant, and resuspending the cells with electrotransfer buffer to give a cell density of 1X10 7 Plasmid pCDNA3.1-GFP, which requires electrotransfection into cells, was added at a concentration of 20ug/ml and gently mixed. The intermittent flow type electrotransfection device is shown in figure 1, the end caps at the two ends of the tube body are respectively sealed by a first electrode and a second electrode, and the first electrode is connected with a hydraulic rod. Before the liquid enters the cavity, a sample injection sheetThe check valve is opened, the sample outlet check valve is closed, the ventilation valve is opened, the air pump is used for pumping the air with the same volume as the pipe body through the ventilation pipe, and the target liquid sample enters the cavity from the liquid inlet pipe; after the cavity is full, the sampling check valve and the ventilation valve are closed. The hydraulic rod pushes the first electrode to move from the non-working position to the working position, so that the liquid in the cavity bears a certain pressure. At this time, an electric pulse is applied to the first electrode and the second electrode, the pulse voltage is 180 volts, the pulse width is 6 milliseconds, the pulse times are 2 times, the pulse interval is 1 second, the electric pulse enables an electric field to be formed in the cavity, so that a cell membrane forms micropores, and target substances in the liquid sample can enter the cells. After the electrotransfection is completed, the hydraulic rod is able to pull the first electrode back from the working position to the non-working position. After the electrotransfection is finished, the sample outlet one-way valve and the ventilation valve are opened, the air pump is used for filling the air with the volume equal to that of the cavity body through the ventilation pipe, and the target liquid sample is discharged from the liquid outlet pipe. The transfected cell suspension was placed in a centrifuge tube, 1000 rpm, and centrifuged for 5 minutes. Removing supernatant, adding OptiCHO culture medium to resuspend cells, inoculating and culturing in triangular conical flask with culture density of 2×10 6 And/ml, then placing on a shaking table for culture, wherein the rotation speed of the shaking table is 125rpm/min, and the culture conditions are as follows: the temperature was 37℃and the carbon dioxide concentration was 5%. After 24 hours, the cells were observed under a fluorescence microscope and examined for electrotransfection efficiency and cell viability using a flow cytometer.
And (3) observation of experimental results:
in experiment five, it can be clearly observed that a small amount of bubbles appear in the cavity after the pulse, while in experiment six the cavity is still full of liquid and no bubbles are generated.
After 24 hours, the experimental five-cell transfection efficiency is detected to be 61% -63%, and the cell survival rate is detected to be 70% -81%; six cells were tested with a transfection efficiency of 83-% > 87% and a cell viability of 81% -84%.
In summary, the pressurizing cavity provided by the invention can improve the electrotransfection efficiency and has obvious effect of reducing the generation of bubbles.
Although the present invention has been described in detail, modifications within the spirit and scope of the invention will be apparent to those of skill in the art. It should be understood that aspects of the invention and portions of the various embodiments and features of the various embodiments described above and/or in the claims below may be combined or interchanged in whole or in part. As will be appreciated by those skilled in the art, in the foregoing description of the various embodiments, those embodiments which refer to another embodiment may be combined with other embodiments as appropriate. Furthermore, those of ordinary skill in the art will appreciate that the foregoing description is by way of example only, and is not intended to limit the invention.
Claims (13)
1. The intermittent flow type electrotransfection device comprises a tube body, a first electrode and a second electrode, and is characterized in that the first electrode and the second electrode are parallel at two ends of the tube body, and a cavity for accommodating a target liquid sample is formed inside the tube body and the first electrode and the second electrode; the pipe body is provided with at least one liquid inlet and outlet and at least one liquid inlet and outlet valve, and the liquid inlet and outlet valve is positioned at the liquid inlet and outlet of the pipe body and controls the opening and closing of the liquid inlet and outlet; the first electrode and/or the second electrode being relatively movable along the tube body; the device further comprises pressurizing means enabling relative movement of the first and/or second electrode along the tube.
2. The intermittent flow electrotransfection device of claim 1 wherein the tube is provided with at least two liquid inlets and outlets.
3. The batch flow electrotransfection device of claim 2 wherein the liquid port is located between the first and second electrodes.
4. The batch flow electrotransfection device of claim 1 wherein the first and/or second electrodes are disposed in an operative position and an inoperative position.
5. The intermittent flow type electrotransfection device according to claim 1, wherein the tube body is provided with a vent tube, and the vent tube is provided with a vent valve.
6. The intermittent flow type electrotransfection device of claim 5, wherein a vent valve is arranged at the joint of the vent pipe and the pipe body.
7. The intermittent flow type electrotransfection device according to claim 1, wherein sealing devices are arranged at the contact positions of the first electrode and the second electrode with the tube body.
8. The batch flow electrotransfection device of claim 7 wherein the sealing means is selected from the group consisting of sealing gaskets, sealing strips, and other means having sealing properties.
9. The batch flow electrotransfection device of claim 8 wherein the tube and the interior of the first and second electrodes form a closed cavity when the liquid inlet and outlet valve and the vent valve are closed.
10. The batch flow electrotransfection device of claim 9 wherein the pressure value inside the closed chamber is 1.0-2.50Mpa after the first and/or second electrodes are moved relatively along the tube when the liquid inlet and outlet valve and vent valve are closed.
11. The intermittent flow type electrotransfection device according to claim 10, wherein the pressure value inside the closed cavity is 1.1-2.0Mpa.
12. The intermittent flow electrotransfection device of claim 1, wherein the pressurizing device includes, but is not limited to, a balloon, a hydraulic rod, a pneumatic rod, a screw pressurizing device, an electromagnetic force pressurizing device.
13. The intermittent flow type electrotransfection device of any one of claims 1 to 12, wherein the second electrode is fixedly arranged at an end orifice of the tube body.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811054671.0A CN110885857B (en) | 2018-09-11 | 2018-09-11 | Intermittent flow type formula electrotransfection device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811054671.0A CN110885857B (en) | 2018-09-11 | 2018-09-11 | Intermittent flow type formula electrotransfection device |
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| Publication Number | Publication Date |
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| CN110885857A CN110885857A (en) | 2020-03-17 |
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| CN205205150U (en) * | 2015-12-01 | 2016-05-04 | 陈剑 | Electric shock pipe |
| CN205205151U (en) * | 2015-12-01 | 2016-05-04 | 陈剑 | Electric shock pipe |
| CN106701565A (en) * | 2015-09-07 | 2017-05-24 | 美天施生物科技有限责任公司 | Disposable cartridge for electroporation |
| CN209098693U (en) * | 2018-09-11 | 2019-07-12 | 苏州壹达生物科技有限公司 | A kind of intermittent streaming electrotransfection device |
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| WO2011142813A1 (en) * | 2010-05-12 | 2011-11-17 | Cellectis Sa | Dynamic mixing and electroporation chamber and system |
| US9382510B2 (en) * | 2011-08-25 | 2016-07-05 | Jian Chen | Methods and devices for electroporation |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106701565A (en) * | 2015-09-07 | 2017-05-24 | 美天施生物科技有限责任公司 | Disposable cartridge for electroporation |
| CN205205150U (en) * | 2015-12-01 | 2016-05-04 | 陈剑 | Electric shock pipe |
| CN205205151U (en) * | 2015-12-01 | 2016-05-04 | 陈剑 | Electric shock pipe |
| CN209098693U (en) * | 2018-09-11 | 2019-07-12 | 苏州壹达生物科技有限公司 | A kind of intermittent streaming electrotransfection device |
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