CN212426065U - Electrical stimulation cell culture device - Google Patents

Abstract

The utility model provides an electrical stimulation cell culture device, which comprises a dish body, a dish cover covered on the upper part of the dish body and a support frame below the dish body, wherein the dish body comprises a dish body inner wall conductive layer and a dish body outer wall power generation layer which are mutually communicated, and the conductive layer and the power generation layer are made of conductive materials; the support frame includes base, supporting part and through connecting rod and base articulated friction unit, the friction unit includes magnetic force district, insulating material district and cup joints with the connecting rod through the through-hole. The culture device has the advantages of low cost, simple operation, no influence on the internal environment of the incubator and the like.

Description

Electrical stimulation cell culture device

Technical Field

The utility model relates to a cell biology technical field especially relates to an electro photoluminescence cell culture apparatus.

Background

Bioelectricity is widely present in the human body and plays an essential role in maintaining normal biological functions. For example, many biological processes including embryogenesis, wound healing, tissue repair and remodeling, and normal growth of organisms are regulated by endogenous bioelectric currents, while neural, muscular, and glandular tissues transmit electrical signals/pulses through internal electric fields. Studies have shown that endogenous electric fields in vivo can regulate biological processes by controlling organ asymmetry through transport of ionic species and macromolecules, membrane voltage, and the axis of cell division. The bioelectric signals enter epigenetic and transcriptional cascades, triggering changes in cellular properties such as proliferation, differentiation, migration, shape changes, gene expression, and apoptosis.

Specifically, biophysical changes in the electric field can be triggered at the cell surface, affecting membrane protein functions such as enzymatic activity (Na +/K + atpase and Ca2+ atpase), membrane receptor complexes, and ion transport channels by altering the charge distribution (i.e., conformation) on biomolecules. Based on the principle, in-vitro simulation of in-vivo bioelectric stimulation is widely applied to excitable cells such as nerve cells, muscle cells, skeletal muscles, heart cells and the like, has important effect on non-excitable cells such as stem cells and osteoblasts, and plays an important role in the field of improving plasmid transfection efficiency.

However, the existing in vitro simulation in vivo biological electrical stimulation culture device mainly depends on a power supply device with stimulation current connected into a culture system, the devices are provided with complex devices such as electrodes, connecting wires, an electric control system, a power supply and the like, the device is integrated into a culture box, and the method has complex engineering and high cost; or the electric control system is externally arranged through a connecting wire, so that the tightness of the incubator is influenced and the operation is complex.

Therefore, the development of the cell culture device which has low cost and simple operation and does not influence the internal environment of the incubator is extremely beneficial to the popularization of the in-vitro electrical stimulation cell culture technology.

SUMMERY OF THE UTILITY MODEL

An object of the utility model is to provide an electro photoluminescence cell culture apparatus, it has low cost, easy operation, does not influence advantages such as incubator internal environment again.

In developing the electrical stimulation cell culture device of the present invention, we considered the source of the electrical stimulation to be of great importance, and we found the electrostatic pulse after studying numerous electrical stimulation sources. The frictional electrification phenomenon is visible everywhere in life, mechanical energy can be converted into electric energy by utilizing the principles of frictional electrification and electrostatic induction, and the electric energy can be used as a power supply. For example, chinese patent publication nos. CN203445807U, CN205051599U, CN104052327A, etc. refer to technologies that convert various forms of mechanical energy into electrical energy through different structural designs. Through reasonable design, the device manufactured based on the principle of triboelectrification and electrostatic induction can be utilized to generate a pulse electric signal with controllable frequency and adjustable output magnitude, and the device can be applied to the technical field of biomedical cytobiology. The method can prevent other materials such as electrodes from contacting cells or cell culture media, greatly reduces the possibility of cell pollution, does not need to be connected with an external controller through a connecting wire, keeps the sealing property and stability of the culture environment, and is beneficial to popularization and application of the electrical stimulation cell culture technology.

Specifically, the technical scheme of the utility model is that: the utility model provides an electrical stimulation cell culture device, includes the ware body, covers at ware lid and the support frame in ware body below on ware body upper portion, and wherein the ware body is including the ware body inner wall conducting layer and the ware body outer wall electricity generation layer of intercommunication each other, and the intercommunication mode can be for built-in wire connection or the intercommunication that extends each other. The conducting layer and the power generation layer are made of conducting materials, and when the conducting materials are rubbed by an insulating material according to a triboelectrification principle, the conducting materials are easy to lose electrons so as to generate an electrostatic effect; the support frame includes base, supporting part and through connecting rod and base articulated friction unit, the friction unit includes magnetic force district, insulating material district and cup joints with the connecting rod through the through-hole.

During the use, whole device all can disinfect, will assemble the culture medium of pouring into and inoculate and wait to cultivate the cell in the ware body of the good and disinfected culture apparatus, cover the ware and put into the incubator after, set up magnetic means in the outside magnetic field of incubator just right region when needing to carry out the electro photoluminescence, based on the principle that repels mutually at the same level, magnetic means's existence makes the magnetic field of friction unit extrude to the ware body, and the insulating material of friction unit pastes the layer of generating electricity. At the moment, the magnetic device is periodically moved back and forth along the direction of the connecting rod, the magnetic zone drives the friction unit to periodically rub back and forth on the power generation layer to generate electrostatic pulses, and the electrostatic pulses are transmitted to cells in the culture medium through the conductive layer to generate a stimulation effect on the cells and influence the cell characteristics such as proliferation, apoptosis, differentiation, membrane protein change and the like. After the electric stimulation is finished, only the external magnetic field needs to be removed, and after the action of the magnetic force is lost, the friction unit is separated from the contact with the power generation layer due to the action of gravity.

Preferably, the conductive layer and the power generation layer are non-ferromagnetic conductive materials, the conductive materials include metal materials, semiconductor materials and conductive polymer materials, the materials can lose electrons and conduct electricity after the insulating materials are rubbed, in order to avoid being attracted by the magnetic force area of the rubbing unit, if the conductive materials are metal materials, ferromagnetic materials such as iron metal and the like are avoided, the non-ferromagnetic metal materials are moderate in cost due to the fact that the conductive efficiency of copper is higher, and the metal copper materials are more preferable.

The dish body can be manufactured in an electroplating mode under the condition of selecting a copper material, and the process is simple and high in efficiency.

Preferably, the friction unit insulating material area is polytetrafluoroethylene nylon material, and the insulating material is the material that does not conduct electricity under the permissible voltage, including nylon, polyformaldehyde, polyoxyethylene, polyimide, polyvinylidene fluoride, polystyrene, polymethyl methacrylate, polyvinyl chloride, polydimethylsiloxane, polytetrafluoroethylene etc. because polytetrafluoroethylene's security is high and the electron power is extremely strong, as preferred scheme, the friction unit passes through viscose and magnetic force district bonding fixation.

Preferably, the power generation layer is located at the bottom of the outer side of the dish body, the friction unit is located below the dish body, the structure is favorable for assembling the device, and the magnetic field is changed below the bottom of the incubator when the device is used.

Preferably, the power generation layer is communicated with the conductive layer through the through hole, the raw materials are less in use in a through hole communication mode, and the cost is lower.

Preferably, the power generation layer is positioned on the outer side wall of the dish body, the friction unit is positioned on one side of the dish body, the structure is relatively easy to operate, the magnetic field changing operation is carried out on the side wall of the incubator corresponding to the magnetic force area, and the operation space is larger and more free.

Preferably, the power generation layer extends inwards from the outer side wall and is communicated with the conducting layer, and the process is simpler in a mode that the power generation layer extends inwards and is communicated with the conducting layer because the outer side of the side wall of the vessel body is a friction electrification region.

Preferably, the magnetic force region is a permanent magnet, such as but not limited to a rare earth permanent magnet material, a ferrite permanent magnet material, an alnico permanent magnet material, etc., and the magnetic force of the permanent magnet is stronger and more obvious.

The support frame is made of any non-ferromagnetic material with a supporting function, i.e. not attracted by an external magnetic field, such as but not limited to an aluminum alloy support, a stainless steel support, various plastic supports, etc., although a non-conductive plastic support is preferably used for reducing the loss of electric charges.

Implement the embodiment of the utility model provides a, following beneficial effect has:

the utility model discloses an electro photoluminescence cell culture device, because simple structure need not to be connected with external control ware through the connecting wire, has kept the seal and the stability of cultivateing the environment, is of value to the popularization and application of electro photoluminescence cell culture technique.

Drawings

FIG. 1 is a schematic view showing the construction of an electrically stimulated cell culture apparatus according to example 1;

FIG. 2 is an exploded view of the parts of an electro-stimulating cell culture apparatus according to example 1;

FIG. 3 is a sectional view of an electrically stimulated cell culture apparatus according to example 1;

FIG. 4 is a schematic view of the structure of an electrically stimulated cell culture apparatus according to example 2;

FIG. 5 is an exploded view of the parts of an electro-stimulating cell culture apparatus according to example 2;

FIG. 6 is a sectional view of an electrically stimulated cell culture apparatus according to example 2;

FIG. 7 is a graph showing the voltage profile of the output of the pulsed electrical stimulation of the electrically stimulated cell culture apparatus described in example 3;

FIG. 8 is a graph showing the current profile of the output of the pulsed electrical stimulation of the electrically stimulated cell culture apparatus described in example 3;

FIG. 9 is a graph showing a comparison of transfection efficiencies of the experimental group and the control group in example 3.

Detailed Description

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.

Example 1:

referring to fig. 1 to 3, the electrical stimulation cell culture device of the present embodiment includes a dish body 01, a dish cover 02 covering the upper portion of the dish body 01, and a support frame 03 below the dish body 01, wherein the dish body 01 includes a conductive layer 011 on the inner wall of the dish body 01 and a power generation layer 012 on the outer wall of the dish body 01, which are communicated with each other, and the conductive layer 011 and the power generation layer 012 are made of conductive materials; wherein, support frame 03 includes base 031, supporting part 032 and through connecting rod 033 and the articulated friction unit 04 of base 031, friction unit 04 includes magnetic field 041, insulating material district 042 and cup joints with connecting rod 033 through-hole 043, and friction unit 04 can slide on connecting rod 033 through the relation of cup jointing.

Further, the conductive layer 011 and the power generation layer 012 are made of non-ferromagnetic conductive material.

Further, the insulating material area 042 of the friction unit 04 is made of teflon.

Further, the power generation layer 012 is located at the bottom of the outer side of the dish body, and the friction unit 04 is located below the dish body 01.

Further, the power generation layer 012 communicates with the conductive layer 011 through the through hole 013.

Further, the magnetic force region 041 is a permanent magnet.

Example 2:

referring to fig. 4-6, the electrical stimulation cell culture device of the present embodiment is different from embodiment 1 in that the electrical generation layer 012 is located on the outer side wall of the dish body 01, and correspondingly, the friction unit 04 is located on one side of the dish body 01, when the embodiment is used, a magnetic field opposite to the magnetic field 041 is applied to the outer side of the incubator corresponding to the friction unit 04, so that the friction unit 04 is pushed towards the side wall of the dish body 01, the insulating material area 042 of the friction unit 04 contacts the electrical generation layer 012, and at this time, the magnetic field outside the incubator, which moves up and down along the direction of the connecting rod 033, drives the friction unit 04 to slide up and down under the magnetic traction action, and further, static electricity is generated on the electrical generation layer through the movement of the friction unit 04.

Further, the power generation layer 012 extends inward from the outer side wall to communicate with the conductive layer 011.

Example 3:

selecting a human non-small cell lung cancer cell line A549 for culturing, adding cells and a culture medium into an experimental group, namely the electrical stimulation culture device shown in example 1, and a control group, namely a common culture dish, culturing and amplifying in an incubator, adding a Green Fluorescent Protein (GFP) eukaryotic expression plasmid packaged by lipo2000 liposome when the cell density reaches 60-80%, adding a periodically variable magnetic field at the bottom of the incubator, adjusting the frequency and the intensity of the variable magnetic field, driving a friction material to generate periodic reciprocating motion by a magnet, rubbing the magnet with the bottom surface of the culture dish, and generating electrical stimulation to the cells in the culture dish by utilizing the friction electrification and electrostatic induction effects; stopping stimulation after 1 hour of electrical stimulation, comparing the change of transfection efficiency at different time points after transfection, and observing that the cell culture dish with the electrical stimulation function can improve the transfection efficiency and shorten the transfection time by detecting the transfection efficiency through the flow cytometry.

Wherein the friction material of the electrical stimulation culture device is a nylon film; the conductive material is a copper film; the magnet is a neodymium iron boron strong magnet, the diameter of the magnet is 10mm, and the thickness of the magnet is 2 mm; the frequency of the changing magnetic field was 3.3Hz, the intensity was 2000Gs, as shown in FIGS. 7-8, and a current with a peak value of 6 microamperes was outputted for a voltage of 130V pulse generated by the culture apparatus during the change of the magnetic field.

As shown in FIG. 9, the transfection efficiencies of the experimental group and the control group were measured at 6 hours, 12 hours, 24 hours, and 48 hours after the culture, respectively, and it was found that the transfection efficiency of the experimental group was significantly higher than that of the control group as the culture time increased.

The experimental effect of the above embodiment proves that the electrical stimulation cell culture device of the utility model can generate stable pulse electrical stimulation by periodically changing the external magnetic field, and on the basis of ensuring the electrical stimulation effect, because no other materials such as electrodes contact the cells or the cell culture medium, the possibility of the contamination of the cells is greatly reduced; meanwhile, the culture environment is not required to be connected with an external controller through a connecting wire, so that the sealing property and the stability of the culture environment are kept. The multiple advantages can greatly help the popularization of the electrical stimulation culture technology.

The foregoing is a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, a plurality of modifications and variations can be made without departing from the principle of the present invention, and these modifications and variations are also considered as the protection scope of the present invention.

Claims (8)

1. An electrical stimulation cell culture device is characterized by comprising a dish body, a dish cover covering the upper part of the dish body and a support frame below the dish body, wherein the dish body comprises a dish body inner wall conducting layer and a dish body outer wall power generation layer which are mutually communicated, and the conducting layer and the power generation layer are made of conducting materials; the support frame includes base, supporting part and through connecting rod and base articulated friction unit, the friction unit includes magnetic force district, insulating material district and cup joints with the connecting rod through the through-hole.

2. The device as claimed in claim 1, wherein the conductive layer and the power generation layer are made of non-ferromagnetic conductive material.

3. The electro-stimulation cell culture device as claimed in claim 1, wherein the friction unit insulating material region is a teflon material.

4. The electrically stimulated cell culture device according to claim 1, wherein the power generation layer is located at the bottom of the outer side of the dish body, and the friction unit is located below the dish body.

5. An electrically stimulated cell culture device according to claim 4, wherein the electricity generating layer is in communication with the conductive layer through the through hole.

6. The electrically stimulated cell culture device according to claim 1, wherein the power generation layer is located on the outer side wall of the dish body, and the friction unit is located on one side of the dish body.

7. An electro-stimulating cell culture device as claimed in claim 6, wherein the electricity generating layer extends inwardly from the outer side wall in communication with the conductive layer.

8. The electro-stimulation cell culture device according to claim 1, wherein the magnetic field is a permanent magnet.

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