KR101637083B1 - Cell culture plate installed on cell culture device - Google Patents

Abstract

It is possible to analyze the culturing process of precise cells of various single cell units and to provide a technique for effectively studying the cultivation of various cells by stably providing mechanical and periodic stimulation as an in vivo environment outside the living body. The cell culture dish installed in the cell culture apparatus according to an embodiment of the present invention has a predetermined pattern portion of the Yin-Yang structure on the upper surface thereof, Wherein the cell culture dish is formed so as to have a pattern having a yin and yang structure corresponding to the pattern portion of the yin and yang structure formed in the cell culture apparatus, and the cell culture dish and the cell A first surface formed so that the culture apparatus can be engaged; And at least one cell immobilization cavity is formed so that the stimulation generated in the cell culture apparatus is delivered to cells to be cultured in a state where the cell culture dish is installed in the cell culture apparatus, And a second surface.

Description

A cell culture plate installed on a cell culture apparatus (cell culture plate installed on cell culture apparatus)

TECHNICAL FIELD [0001] The present invention relates to a cell culture dish installed in a cell culture apparatus capable of efficiently culturing cells and living tissues by mechanical stimulation, more specifically, to provide cells with static and dynamic loads similar to those in a living body, The present invention relates to a cell culture dish capable of being installed in a cell culture apparatus so as to efficiently transmit mechanical stimulation of a culture apparatus and simultaneously cultivate various cells in a stable state.

In general, for the study of cells and tissues, the cells to be cultured are placed in a constant culture apparatus, and a process of stimulating or natural culturing is analyzed.

Generally, in order to cultivate cells and tissues, the cells to be cultured are placed on a cell culture dish composed of hard plastic or glass, and are cultured in a static and passive manner.

However, in vivo environments where cell tissue is actually active, the cells are mechanically very dynamic stimuli. Therefore, in order to study the culture condition of cells effectively, it is necessary to experimentally investigate how such mechanical stimulation affects the behavior and growth of cells and tissues. However, in the present cell culture system, It is difficult.

In order to overcome this problem, a technique of applying various mechanical stimuli to cells outside of the living body has been used. For example, cells are cultured by applying a negative pressure in a micropipette, applying a mechanical stimulus by magnetic beads and magnets, applying a mechanical stimulus by introducing fluid, or applying an external force to relax and shrink the flexible plate have.

However, these devices are usually difficult to analyze and observe the differentiation and culturing of single cells in a unit of centimeter area, and thus it is difficult to analyze the precise differentiation process of cells.

In addition, when various cells are cultured in the same apparatus at the same time, it is an optimal culture environment for each cell to provide an environment in which various stimuli and cells can be fixed according to the size of the cells. It is recognized as an essential task.

Accordingly, it is an object of the present invention to provide a method and apparatus for analyzing the culturing process of precise cells of various single cell units and stably providing mechanical and periodic stimulation as an in-vivo environment outside the living body, The purpose is to provide technology.

In order to achieve the above object, a cell culture dish installed in a cell culture apparatus according to an embodiment of the present invention includes a layer comprising a predetermined pattern portion of a Yin-Yang structure on an upper surface thereof, a layer comprising an electroactive polymer material, A cell culture dish provided in a cell culture apparatus including an electrode portion coated on one region of the layer, wherein the cell culture dish has a pattern having a yin-yang structure corresponding to a pattern portion of the yin and yang structure formed in the cell culture apparatus, A first surface formed to have the cell culture dish and the cell culture device fitted to each other; And at least one cell immobilization cavity for allowing the stimulation generated in the cell culture apparatus to be delivered to cells to be cultured in a state where the cell culture dish is installed in the cell culture apparatus, And a second surface on which the first electrode is formed.

According to the present invention, a cell culture apparatus composed of an electroactive polymer transmits mechanical stimulation when a voltage is applied to an electroactive polymer to the cell culture dish of the present invention, Lt; RTI ID = 0.0 > cell < / RTI >

In addition, it is possible to prevent the direct application of an electrical load to the cell culture apparatus, thereby ensuring the stability of the cell culture process.

In addition, it is possible to cultivate a plurality of cells simultaneously in one cell culture apparatus through a cavity having various rigidities and concavo-convex structures, and thus it is possible to rapidly and efficiently cultivate healthy cells.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a side cross-sectional view showing a configuration of a cell culture apparatus provided with a cell culture dish according to an embodiment of the present invention; FIG.
Figures 2 and 3 are side cross-sectional and plan views of a cell culture dish according to one embodiment of the present invention.
4 and 5 are side cross-sectional views of a cell culture apparatus and a cell culture dish in a state where cells are placed according to an embodiment of the present invention.

Hereinafter, a cell culture apparatus according to each embodiment of the present invention will be described with reference to the accompanying drawings. 1 to 5 referred to in the following description, the layer, the pattern part and the thickness, size and shape of the cell are configured to facilitate identification for the purpose of explanation of the present invention, but actually, the layer, the pattern part, It is natural that it is composed of a micro structure.

1 is a side cross-sectional view showing a configuration of a cell culture apparatus in which a cell culture dish according to an embodiment of the present invention is installed.

1, a cell culture apparatus 10 in which a cell culture dish according to an embodiment of the present invention is installed includes a layer 11 including an electroactive polymer material, And an electrode (14) applied to the electrode.

Particularly, the layer 11 is characterized in that a pattern portion 15 of a negative / negative structure (or concave-convex structure) is formed on the upper surface, for example, on the side where the cell culture dish is located.

It is preferable that the layer 11 of the cell culture apparatus 10 maintains a state in which the layer 11 is subjected to a force to improve efficient mechanical stimulation and enhance stability to electricity. To this end, there is a fixing part 16 for fixing one end of the layer 11 in the extending direction, and a supporting part 13 for supporting the layer 11 may be included, (11) can be fixed by the fixing means (12). Of course, it goes without saying that any configuration capable of maintaining the layer 11 in a stretched state can be included other than the above examples.

In a region where the electrode portion 14 is applied, when voltage is applied, mechanical deformation occurs due to the characteristics of the electroactive polymer material, thereby providing a mechanical stimulus similar to the inside of the body to the cells.

The electroactive polymer material is deformed in a ratio of several tens of percent when electrical stimulation is applied. At this time, the portion to which the electrode is applied is compressed in the thickness direction by the Maxwell force, and since the material is incompressible, it exhibits a stretching behavior in the plane direction correspondingly.

When the electroactive polymer material is applied to the present cell culture apparatus as the layer 11, it can be configured to have a very thin (micrometer-unit) thickness. Accordingly, if the layer 11 is not supported, a flat state can not be maintained due to gravity in a state where a voltage is not applied, and the layer 11 is bent downward, and the cells are not properly supported.

Further, in order to further increase the strain of the electroactive polymer material and to expect stable and stable behavior, it is necessary to maintain the electroactive polymer material in a stretched state beforehand.

One end of the layer 11 is fixed to the fixing portion 16 and the supporting portion 13 is fixed to the fixing portion 16 while supporting the layer 11 And the other end of the layer 11 is combined with the layer 11 to fix the layer 11 in a stretched state.

Among the electroactive polymer materials, the electrode part 14 to which the electrode is applied is deformed as voltage is applied as mentioned above. Therefore, when the amplitude and the frequency of the voltage applied to the layer 11 are controlled, periodic stimulation can be applied, and the rate at which the thickness and the width of the layer 11 are deformed and the period of deformation can be easily controlled, It is possible to provide a customized cell culture apparatus according to the size and type of cells by forming a cell-friendly environment in which complicated and various biological mechanisms are simulated. To this end, a processor (not shown) may be installed in the cell culture apparatus 10 to control the amplitude and the frequency of the voltage applied to the electrode unit 14.

Further, in various embodiments of the present invention, the shape of the layer, the stretching direction, the shape of the fixing portion, the shape of the electrode, the size and the pattern, etc. may be variously configured and accordingly include a configuration suitable for culturing various cells You can.

The description of the cell culture dish in which the culture medium and the cells are placed by being installed in the cell culture apparatus 10 mentioned above is mentioned in FIGS. 2 and 3. FIG.

2 and 3 are side sectional views and plan views of a cell culture dish according to an embodiment of the present invention.

Referring to FIG. 2, the cell culture dish 100 according to an embodiment of the present invention includes a first surface 120 and a second surface 110.

The first surface 120 is formed to have a pattern having a minus-yang structure corresponding to the above-mentioned pattern portion 15 formed in the cell culture apparatus 10 of Fig. 1, so that the cell culture dish 100 and the cell culture apparatus (10) to be fitted to each other.

On the other hand, the second surface 110 is formed in the cell culture apparatus 10 in such a manner that the cell culture dish 100 is mounted on the cell culture apparatus 10 by the structure of the first surface 120, Characterized in that at least one cell immobilization cavity is formed so that the cell can be fixed and positioned while allowing the stimulation to be delivered to cells to be cultured.

In the example shown in FIG. 2, the cell fixing cavity means a concavo-convex structure formed on the second surface 110. Referring to FIG. 3, it can be confirmed that the cavity 101 of the concave-convex structure is formed in the cell culture dish 100. 4, the cell immobilization cavity may have a planar shape other than the same shape as that of the second surface 110 of FIG. 2, and a structure in which at least one cell can be positioned at the same time have.

The cell culture dish 100 may be made entirely of an electrical insulator. The cell culture apparatus 10 is composed of an electroactive polymer material, and is mechanically deformed by application of a voltage. At this time, since the cells can be necrosis by electricity, the cell culture dish 100 can be formed of an electric insulator to prevent necrosis of the cells.

In order to transmit the active deformation amount of the cell culture apparatus 10 as much as possible to the cell culture dish 100 and to effectively transmit the three-dimensional mechanical deformation to the cells while minimizing the amount of active deformation, 100) should have elasticity and flexibility. Further, it should be easy to treat a base (for example, collagen) for attaching cells to the cell culture dish 100.

In order to perform the above function, the cell culture dish 100 may be made of a polymer material such as a PDMS (Polydimethylsiloxane) material. Of course, other substances or mixtures of substances other than PDMS may be used as long as they are capable of performing the above functions of the cell culture dish 100 .

The cell culture dish 100 may be bonded to the cell culture apparatus 10 by the above-described structure, but other types of bonding structures may be formed to increase the binding force.

For example, the cell culture dish 100 and the cell culture apparatus 10 may be attached to the first surface 120 by a thermal compression process at a temperature lower than the melting point of the material constituting the cell culture dish 100, Lt; / RTI > Alternatively, the cell culture dish 100 and the cell culture apparatus 10 may be combined using the same material as the material constituting the cell culture dish 100 as an adhesive.

For example, the PDMS may be thermally pressed in a range of 185 to 195 degrees Celsius, which is 10 to 20 degrees Celsius lower than the melting point of PDMS, or the PDMS itself may be used as an adhesive.

When using PDMS, the cell culture dish 100 can be manufactured by using a device capable of spraying and applying a PDMS solution mixed with a solvent onto a mold having a concave-convex structure by a method such as ultrasonic spraying.

At this time, the rigidity and structure of the cell culture dish 100 can be diversified so that a plurality of cells can be simultaneously cultured in one cell culture dish 100. An example of this is shown in Figs. 4 and 5 are examples of side sectional views of a cell culture apparatus and a cell culture dish in a state where cells are placed according to an embodiment of the present invention.

First, referring to FIG. 4, it can be confirmed that the cell culture dish 100 is installed in the cell culture apparatus 10. At this time, the culture liquid 106 may be present in the cavity in which the cells 20 of the cell culture dish 100 are raised, and the cells 20 are arranged in one region of the cavity by the culture liquid 106.

At this time, it can be confirmed that the cavity in which the cells 20 are fixed is divided by the first to third regions 103, 104, and 105.

This means that reinforcing powders having different ratios are added to the regions 103, 104, and 105 so that the cavities of the culture dish can have various rigidities.

When the reinforcing powder is added at different ratios, the regions 103, 104, and 105 will have different degrees of deformation due to the mechanical deformation of the cell culture apparatus 10, The stimulus also changes.

That is, since the degree of mechanical stimulation transmitted to the cell culture dish 100 from the cell culture apparatus 10 is varied by the addition of the reinforcing powder, the addition rates of the reinforcing powders are different for each of the regions 103, 104, and 105 , The cell culture environment in each of the regions 103, 104, and 105 can be controlled differently by one cell culture apparatus 10, and the effect of enabling the simultaneous cultivation of a plurality of cells 20 can be achieved.

4, the outer wall 102 may be formed at the outermost part of the cell culture dish 10. [ Reinforced powder is added to the outer wall 102 to improve the structural stability of the cell culture dish 100 and to prevent the outflow of the culture liquid 106. [

As the reinforcing powder, for example, graphene, CNT, nano-clay, hydroxyapatite and the like may be used, but the present invention is not limited thereto.

5, in the state where the cell culture dish 100 is installed in the cell culture apparatus 10 and the cells (no identification number) are arranged inside the outer wall 102 and the culture liquid 106 as in FIG. 4, .

At this time, in FIG. 5, the fourth to sixth regions 107, 108, and 109 can be identified. As can be seen from FIG. 5, the fourth to sixth regions 107, 108, and 109 have different cavity sizes.

In Fig. 4, reinforcing powder is added to the cell culture dish 10 at different ratios to have different stiffness, so that the mechanical stimuli delivered to the cells 20 by the same cell culture apparatus 10 can be different from each other .

On the other hand, in FIG. 5, different sizes of cavities formed in the fourth to sixth regions 107, 108, and 109 can allow cells of different sizes to be simultaneously cultured by one cell culture apparatus 10.

Of course, the embodiments of FIGS. 4 and 5 may be implemented in a state where they are not independent of each other.

With this configuration, various cells can be cultured in a stable environment at the same time, enabling healthy cell culture to be performed quickly and efficiently, and a positive effect can be provided in expanding application of cell and tissue culture technology.

While the present invention has been described in connection with what is presently considered to be the most practical and preferred embodiments, it is to be understood that the invention is not limited to the disclosed embodiments. That is, within the scope of the present invention, all of the components may be selectively coupled to at least one.

It is also to be understood that the terms such as " comprises, "" comprising," or "having ", as used herein, mean that a component can be implanted unless specifically stated to the contrary. But should be construed as including other elements. All terms, including technical and scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, unless otherwise defined. Commonly used terms, such as predefined terms, should be interpreted to be consistent with the contextual meanings of the related art, and are not to be construed as ideal or overly formal, unless expressly defined to the contrary.

The foregoing description is merely illustrative of the technical idea of the present invention and various changes and modifications may be made by those skilled in the art without departing from the essential characteristics of the present invention. Therefore, the embodiments disclosed in the present invention are intended to illustrate rather than limit the scope of the present invention, and the scope of the technical idea of the present invention is not limited by these embodiments. The scope of protection of the present invention should be construed according to the following claims, and all technical ideas within the scope of equivalents should be construed as falling within the scope of the present invention.

Claims (7)

1. A cell culture dish provided in a cell culture apparatus comprising a predetermined pattern portion of a positive and negative structure on an upper surface thereof, a layer including an electroactive polymer material, and an electrode portion coated on one region of the layer,
The cell culture dish was prepared by dissolving,
A first surface formed to have a pattern having a minus-odd structure corresponding to a pattern part of a minus-and-heath structure formed in the cell culture apparatus, the cell culture dish and the cell culture apparatus being fitted to each other; And
Wherein the cell culture dish is installed in the cell culture apparatus so that the stimulation generated in the cell culture apparatus is delivered to the cells to be cultured and at least one cell immobilization cavity And a second surface formed on the surface of the cell culture dish. The method according to claim 1,
Wherein the at least one cell-fixing cavity formed on the second surface comprises:
Wherein a cavity having a different size is formed in the cell culture apparatus. The method according to claim 1,
The cell culture dish was prepared by dissolving,
A cell culture dish provided in a cell culture apparatus characterized by comprising an electric insulator. The method according to claim 1,
The cell culture dish was prepared by dissolving,
Wherein the cell culture dish is installed in a cell culture apparatus. The method according to claim 1,
The cell culture dish was prepared by dissolving,
Wherein a reinforcing powder having a different ratio is added to each of the regions so as to have different stiffnesses for the respective regions. The method according to claim 1,
Wherein an outer wall is formed at the outermost part of the cell culture dish, and a reinforcing powder is added to the outer wall of the cell culture dish. The method according to claim 1,
The cell culture apparatus and the cell culture dish are,
At least one of a thermocompression process at a temperature lower than a melting point of a material constituting the cell culture dish and a process of adhering the cell cultivation dish using the same material as the material constituting the cell cultivation dish as an adhesive Wherein the cell culture dish is attached to the cell culture apparatus by one process.

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